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BITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
10 TIPS FOR CONSISTENT QPCR Quantitative PCR (qPCR) uses fluorescent dyes or probes to visualize the amplification of specific DNA sequences as it happens (i.e. in real time). The dyes or probes fluoresce when they bind to newly amplified DNA, and the amount of fluorescence emitted is proportional to the amount of DNA (or mRNA) present in the original sample. By detecting newly synthesized DNA during the exponential ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
THE BOOM METHOD FOR NUCLEIC ACID PURIFICATION: THE The Boom method, or Boom nucleic acid extraction method, is a solid phase extraction technique for isolating nucleic acids from a solution of biological matter. This is just a fancy way of saying you use this technique to expose and remove the nucleic acids from a cell. First developed by William R. Boom, the Boom method makes use of silica-coated magnetic beads (SiO2) that bind HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, and PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in USING A GEL FILTRATION CHROMATOGRAM TO ESTIMATE MOLECULAR Gel filtration chromatography (also known as size exclusion chromatography, molecular sieve chromatography, or gel permeation chromatography) is based on the differential distribution of the components in a sample between the mobile and stationary phases. Specifically, in gel filtration chromatography, this differential distribution depends on the size and shape of the components.BITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
10 TIPS FOR CONSISTENT QPCR Quantitative PCR (qPCR) uses fluorescent dyes or probes to visualize the amplification of specific DNA sequences as it happens (i.e. in real time). The dyes or probes fluoresce when they bind to newly amplified DNA, and the amount of fluorescence emitted is proportional to the amount of DNA (or mRNA) present in the original sample. By detecting newly synthesized DNA during the exponential ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
THE BOOM METHOD FOR NUCLEIC ACID PURIFICATION: THE The Boom method, or Boom nucleic acid extraction method, is a solid phase extraction technique for isolating nucleic acids from a solution of biological matter. This is just a fancy way of saying you use this technique to expose and remove the nucleic acids from a cell. First developed by William R. Boom, the Boom method makes use of silica-coated magnetic beads (SiO2) that bind HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, and PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in USING A GEL FILTRATION CHROMATOGRAM TO ESTIMATE MOLECULAR Gel filtration chromatography (also known as size exclusion chromatography, molecular sieve chromatography, or gel permeation chromatography) is based on the differential distribution of the components in a sample between the mobile and stationary phases. Specifically, in gel filtration chromatography, this differential distribution depends on the size and shape of the components.BITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? SARS-COV-2 VARIANTS ARE ESCAPING NEUTRALIZING ANTIBODY SARS-CoV-2 variants are escaping neutralizing antibody responses, but have not escaped the T-cell response - A Johnson & Johnson Case Study Date: Thursday, June 18th 2021Time: 8.30am Los Angeles, 10.30am Chicago, 11.30am New York, 4.30pm London, 5.30pm Berlin To understand immune evasion of emerging variants and vaccine responses, researchers need to go beyond the antibody response and PAGE 10 – BITESIZE BIO No matter the ingenuity of your science or the capabilities of your core facility, your results are only as good as your peer-to-peer relationship with the flow cytometry personnel. RICHARD HENDERSON (UNIVERSITY OF CAMBRIDGE) #22. In this episode of The Microscopists, we’re joined by molecular biologist, biophysicist and Nobel Laureate Richard Henderson from the MRC Laboratory of Molecular Biology, Cambridge. We’ll discuss some of his earlier career challenges in biophysics and what attracted him to biology, his pioneering work in the field of electron microscopy, his favorite comic books, and passion for ARE YOU READY FOR YOUR FIRST QPCR? Hope you had a heavy breakfast, because the first qPCR is going to take time. Did you remember to fill your coffee cup? There are a lot of intricacies in qPCR that will need your neural networks to be brisk. How qPCR differs from traditional PCR Unlike traditional PCR, qPCR measures the amplification of DNA in real-time. As the reaction proceeds, a graph is plotted by the software with PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
PROBLEMS AMPLIFYING GC-RICH REGIONS? PROBLEM SOLVED! The Cause of Your Problems. Problem 1. Stability: Thermal and Structural. GC-rich DNA sequences are more stable than sequences with low GC-content. For PCR, this means that the higher the GC content, the higher the melting point of the DNA. Under pressure, such as when exposed to heat, the GC-rich sequences can take far more abuse thanGC-low
PRACTICAL APPLICATION OF PHENOL/CHLOROFORM EXTRACTION While there are many more methods to choose from for cleaning up your RNA or DNA than there used to be, sometimes Phenol/Chloroform extraction is still the best way to go. Here I’ll discuss some of the practical aspects of using this technique. Nick introduced the topic of Phenol/Chloroform extraction in a previous article, touching on some of the ideas about how organic extraction will USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed SELECTING CELL LINES: ENSURE YOU CHOOSE THE RIGHT ONES FOR One of the most rapidly-dividing cell lines is EB66, which doubles every 12-14 hours. Insect cells like Sf9 and Sf21 double every 24 hours or so, comparable with HeLa (about 23 hours). Don’t be fooled into thinking that all human cell lines grow at a similar rate – HEK293 double every 36 hours, but CACO2 take around 80 hours. CPEC– A QUICK AND INEXPENSIVE CLONING STRATEGY CPEC is a sequence-independent cloning strategy, making it easy to assemble multiple fragments into any vector and carry out complex library preparations (unlike sequence dependent cloning like Gateway or Univector). It is also flexible as to its use unlike traditional cloning or TA cloning that is restricted in application. HOW AND WHY TO USE FEEDER CELLS WHEN CULTURING PRIMARY CELLS Feeder cells help in the long-term establishment of difficult cultures while maintaining their stemness and regenerative capability. Although feeder-free systems for primary cell cultures have been recently developed, many hESC, hiPSC, epidermal and other epithelial stem cells have shown their dependency on feeder cells to grow and proliferate. ANTIBIOTIC DISPOSAL IN THE LAB: SIMPLE TIPS TO GET IT RIGHT Although this article provides basic information on the proper disposal of antibiotics, it is imperative to consult and follow institutional guidelines to ensure the correct disposal of your laboratory waste – as rules may differ by location. Finally, it is crucial to have careful and responsible practice for safe disposal ofantibiotic waste
TROUBLESHOOTING AGAROSE GELS: TOP TIPS FOR PERFECT IMAGES Let the Gel Take Its Time. This is the most important tip: don’t rush an agarose gel! You don’t want your beautiful cloning result to be ruined by a terrible picture just because you were in a hurry. You have to run your gel at under 75V and make sure the buffer is not overheating. You can reduce the risk of overheating by cooling downthe
SERUM IN CELL CULTURE: SHOULD YOU USE IT AND HOW CAN YOU STOP? And you will often add serum, such as FBS. This is one of the main ingredients in cell culture media because it contains hormones, lipids and growth factors essential for cell proliferation and growth. If you remove the serum from your culture medium, you will notice a big difference. Deliberate serum-starvation is used to induce cell cycle HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed SELECTING CELL LINES: ENSURE YOU CHOOSE THE RIGHT ONES FOR One of the most rapidly-dividing cell lines is EB66, which doubles every 12-14 hours. Insect cells like Sf9 and Sf21 double every 24 hours or so, comparable with HeLa (about 23 hours). Don’t be fooled into thinking that all human cell lines grow at a similar rate – HEK293 double every 36 hours, but CACO2 take around 80 hours. CPEC– A QUICK AND INEXPENSIVE CLONING STRATEGY CPEC is a sequence-independent cloning strategy, making it easy to assemble multiple fragments into any vector and carry out complex library preparations (unlike sequence dependent cloning like Gateway or Univector). It is also flexible as to its use unlike traditional cloning or TA cloning that is restricted in application. HOW AND WHY TO USE FEEDER CELLS WHEN CULTURING PRIMARY CELLS Feeder cells help in the long-term establishment of difficult cultures while maintaining their stemness and regenerative capability. Although feeder-free systems for primary cell cultures have been recently developed, many hESC, hiPSC, epidermal and other epithelial stem cells have shown their dependency on feeder cells to grow and proliferate. ANTIBIOTIC DISPOSAL IN THE LAB: SIMPLE TIPS TO GET IT RIGHT Although this article provides basic information on the proper disposal of antibiotics, it is imperative to consult and follow institutional guidelines to ensure the correct disposal of your laboratory waste – as rules may differ by location. Finally, it is crucial to have careful and responsible practice for safe disposal ofantibiotic waste
TROUBLESHOOTING AGAROSE GELS: TOP TIPS FOR PERFECT IMAGES Let the Gel Take Its Time. This is the most important tip: don’t rush an agarose gel! You don’t want your beautiful cloning result to be ruined by a terrible picture just because you were in a hurry. You have to run your gel at under 75V and make sure the buffer is not overheating. You can reduce the risk of overheating by cooling downthe
SERUM IN CELL CULTURE: SHOULD YOU USE IT AND HOW CAN YOU STOP? And you will often add serum, such as FBS. This is one of the main ingredients in cell culture media because it contains hormones, lipids and growth factors essential for cell proliferation and growth. If you remove the serum from your culture medium, you will notice a big difference. Deliberate serum-starvation is used to induce cell cycle HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water SARS-COV-2 VARIANTS ARE ESCAPING NEUTRALIZING ANTIBODY SARS-CoV-2 variants are escaping neutralizing antibody responses, but have not escaped the T-cell response - A Johnson & Johnson Case Study Date: Thursday, June 18th 2021Time: 8.30am Los Angeles, 10.30am Chicago, 11.30am New York, 4.30pm London, 5.30pm Berlin To understand immune evasion of emerging variants and vaccine responses, researchers need to go beyond the antibody response and INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING As is sadly the case in many experiments, site-directed mutagenesis (SDM) does not always work the way we would like it to the first time around. Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. RICHARD HENDERSON (UNIVERSITY OF CAMBRIDGE) #22. In this episode of The Microscopists, we’re joined by molecular biologist, biophysicist and Nobel Laureate Richard Henderson from the MRC Laboratory of Molecular Biology, Cambridge. We’ll discuss some of his earlier career challenges in biophysics and what attracted him to biology, his pioneering work in the field of electron microscopy, his favorite comic books, and passion for PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. THE CS&T REPORT: ITS TROUBLES, AND HOW TO FIX THEM Laser alignment issue. - Clean with 5min clean solution and 5 min of water can help but long monthly clean is better. - Prime. - Check if the short pass filter is in the right position: the numbers should face inwards. - Align the lasers - engineer might be needed. All the values of one laser are off. PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UP CRISPR-Cas9 editing is a relatively inexpensive way of deleting, silencing, or otherwise modifying a gene or region. If you’re lucky, you can pick up Cas9 and guide RNA expression vectors from a colleague or collaborator’s lab; then all you need to buy are your primers to synthesize the guide RNA vectors. The remaining preparatory steps can THE BOOM METHOD FOR NUCLEIC ACID PURIFICATION: THE The Boom method, or Boom nucleic acid extraction method, is a solid phase extraction technique for isolating nucleic acids from a solution of biological matter. This is just a fancy way of saying you use this technique to expose and remove the nucleic acids from a cell. First developed by William R. Boom, the Boom method makes use of silica-coated magnetic beads (SiO2) that bind FIVE METHODS FOR ASSESSING PROTEIN PURITY AND QUALITY If you’ve ever worked with proteins in the lab, you probably know just how critical protein purity and quality are for downstream applications. In this article, we’ll review the multitude of problems that are encountered with ‘bad’ protein samples and how you can analyze the purity and integrity of your favorite protein prior to using it for important experiments. HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODSSEE MORE ONBITESIZEBIO.COM
HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic 10 TIPS FOR CONSISTENT QPCR Quantitative PCR (qPCR) uses fluorescent dyes or probes to visualize the amplification of specific DNA sequences as it happens (i.e. in real time). The dyes or probes fluoresce when they bind to newly amplified DNA, and the amount of fluorescence emitted is proportional to the amount of DNA (or mRNA) present in the original sample. By detecting newly synthesized DNA during the exponential SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, and USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODSSEE MORE ONBITESIZEBIO.COM
HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic 10 TIPS FOR CONSISTENT QPCR Quantitative PCR (qPCR) uses fluorescent dyes or probes to visualize the amplification of specific DNA sequences as it happens (i.e. in real time). The dyes or probes fluoresce when they bind to newly amplified DNA, and the amount of fluorescence emitted is proportional to the amount of DNA (or mRNA) present in the original sample. By detecting newly synthesized DNA during the exponential SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, andBITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? SARS-COV-2 VARIANTS ARE ESCAPING NEUTRALIZING ANTIBODY SARS-CoV-2 variants are escaping neutralizing antibody responses, but have not escaped the T-cell response - A Johnson & Johnson Case Study Date: Thursday, June 18th 2021Time: 8.30am Los Angeles, 10.30am Chicago, 11.30am New York, 4.30pm London, 5.30pm Berlin To understand immune evasion of emerging variants and vaccine responses, researchers need to go beyond the antibody response and RICHARD HENDERSON (UNIVERSITY OF CAMBRIDGE) #22. In this episode of The Microscopists, we’re joined by molecular biologist, biophysicist and Nobel Laureate Richard Henderson from the MRC Laboratory of Molecular Biology, Cambridge. We’ll discuss some of his earlier career challenges in biophysics and what attracted him to biology, his pioneering work in the field of electron microscopy, his favorite comic books, and passion for A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODS ELISA-based methods can be set up for microbial detection (usually within diagnostics) on a species-by-species basis. These methods are highly sensitive but they rely on very specific antibodies and highly discriminating protein (s) within the organism of interest. 9. Chemical/Analytical Identification. INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. THE BEGINNER’S GUIDE TO READING PLASMID MAPS Let’s start with a classic plasmid: pBR322 1. It is often used as a backbone for derivative vectors because it has all features needed for a successful cloning (Figure 1). As you see from the map center, the size of the linearized plasmid is 4361 base pairs. Before you start working with any plasmid, it is advisable to linearize it by cutting PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in HOMEMADE PCR TEST FOR MYCOPLASMA CONTAMINATION Mycoplasma is one of the biggest threats to cell culture - feared by most researchers who do such experiments. Mycoplasma contamination is relatively common and can persist for a long time while remaining undetected. Typically, one infected culture can spread to other cultures when improper working techniques are used. Besides proper safety measures (putting new cell lines into quarantines PRACTICAL APPLICATION OF PHENOL/CHLOROFORM EXTRACTION While there are many more methods to choose from for cleaning up your RNA or DNA than there used to be, sometimes Phenol/Chloroform extraction is still the best way to go. Here I’ll discuss some of the practical aspects of using this technique. Nick introduced the topic of Phenol/Chloroform extraction in a previous article, touching on some of the ideas about how organic extraction willBITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODSSEE MORE ONBITESIZEBIO.COM
USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, andBITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODSSEE MORE ONBITESIZEBIO.COM
USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
PLASMID DNA ON AGAROSE GEL: THE SECRET OF THE 3 BANDS One Molecule, Many Forms: Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands. When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA will be supercoiled, but other forms can also crop up. How these forms will show up on an agarose gel (in HOW TO FIX ADHERENT CELLS FOR MICROSCOPY AND IMAGING Many cell lines commonly used in research are adherent, meaning they attach and grow on a surface rather than just hanging out in suspension. If you wish to perform imaging of adherent cells, such as to undertake cancer cell microscopy, you need to fix them to your microscope slides. Luckily for you- this is fairly easy to do. Thanks to the fact that adherent cells are, well, adherent, andBITESIZE BIO
17 Ways to Stop Pipetting Errors From Ruining Your Experiments Pipetting errors getting you down? Learn how to avoid and correct errors that hurt your pipetting accuracy in this easy-to-follow article. By Biotix Taming the Literature 18+ Ways to Improve your PubMed Searches Are you getting the most out of your PubMed searches or are you wasting lots of time slogging through pages of results? A QUICK GUIDE TO IDENTIFYING MICROBES: 9 EASY METHODS ELISA-based methods can be set up for microbial detection (usually within diagnostics) on a species-by-species basis. These methods are highly sensitive but they rely on very specific antibodies and highly discriminating protein (s) within the organism of interest. 9. Chemical/Analytical Identification. INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy RICHARD HENDERSON (UNIVERSITY OF CAMBRIDGE) #22. In this episode of The Microscopists, we’re joined by molecular biologist, biophysicist and Nobel Laureate Richard Henderson from the MRC Laboratory of Molecular Biology, Cambridge. We’ll discuss some of his earlier career challenges in biophysics and what attracted him to biology, his pioneering work in the field of electron microscopy, his favorite comic books, and passion for PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. THE BEGINNER’S GUIDE TO READING PLASMID MAPS Let’s start with a classic plasmid: pBR322 1. It is often used as a backbone for derivative vectors because it has all features needed for a successful cloning (Figure 1). As you see from the map center, the size of the linearized plasmid is 4361 base pairs. Before you start working with any plasmid, it is advisable to linearize it by cutting HOW TO PRESERVE MICROORGANISMS: THE LONG (AND SHORT) GAME Given the importance of microorganisms as cell factories in biotechnological applications and as model organisms for the study of various biological processes, the preservation of microorganisms plays a key role in ensuring reproducible results and continuity in research. Maintaining a library of microbial stocks also enables microorganisms to be easily stored and retrieved, as THE PROPER WAY TO USE THE SUB-G1 ASSAY The Proper Way To Use The Sub-G1 Assay. One of the most widely used assays to determine apoptosis by flow cytometry is the estimation of fractional DNA content (aka sub-G1 assay). During apoptosis, genomic DNA is cleaved into smaller fragments, each approximately 180 bp (and multiples of it). This is a specific marker of apoptosis and therefore HOW MEASUREMENT OF CONCENTRATION AND PURITY OF NUCLEIC High quality DNA should have an A 260 /A 280 ratio of 1.7 to 2.0. Other possible contaminants are salt or phenol, which are measured at 230nm. The A 260 /A 230 ratio should be greater than 1.5. So with one sample, you can measure the absorbance at 230, 260 and 280nm to determine both concentration and purity of your nucleic acids. PROBLEMS AMPLIFYING GC-RICH REGIONS? PROBLEM SOLVED! The Cause of Your Problems. Problem 1. Stability: Thermal and Structural. GC-rich DNA sequences are more stable than sequences with low GC-content. For PCR, this means that the higher the GC content, the higher the melting point of the DNA. Under pressure, such as when exposed to heat, the GC-rich sequences can take far more abuse thanGC-low
USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING As is sadly the case in many experiments, site-directed mutagenesis (SDM) does not always work the way we would like it to the first time around. Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT MICROBES ARE USED IN BIOTECHNOLOGY AND HOW? Biotechnology is the use of biological organisms in technological processes. It is almost as old as the civilization itself, although it wasn't called 'biotechnology' until the 20th century. Far from abandoning it in the 21st century, we are developing new uses for biological organisms. 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
CPEC– A QUICK AND INEXPENSIVE CLONING STRATEGY CPEC is a sequence-independent cloning strategy, making it easy to assemble multiple fragments into any vector and carry out complex library preparations (unlike sequence dependent cloning like Gateway or Univector). It is also flexible as to its use unlike traditional cloning or TA cloning that is restricted in application. SELECTING CELL LINES: ENSURE YOU CHOOSE THE RIGHT ONES FOR One of the most rapidly-dividing cell lines is EB66, which doubles every 12-14 hours. Insect cells like Sf9 and Sf21 double every 24 hours or so, comparable with HeLa (about 23 hours). Don’t be fooled into thinking that all human cell lines grow at a similar rate – HEK293 double every 36 hours, but CACO2 take around 80 hours. THE CS&T REPORT: ITS TROUBLES, AND HOW TO FIX THEM Laser alignment issue. - Clean with 5min clean solution and 5 min of water can help but long monthly clean is better. - Prime. - Check if the short pass filter is in the right position: the numbers should face inwards. - Align the lasers - engineer might be needed. All the values of one laser are off. HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING As is sadly the case in many experiments, site-directed mutagenesis (SDM) does not always work the way we would like it to the first time around. Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. HOW CELL-FREE SYSTEMS WORK AND WHEN YOU SHOULD USE THEM Cell-free expression systems, which take expression machinery out of the cell and into a test tube, are a solution to this seemingly intractable problem. These systems can be used to answer fundamental biological questions, such as studying expression in “protocells”. They are equally useful for quickly prototyping genetic and metabolic CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT MICROBES ARE USED IN BIOTECHNOLOGY AND HOW? Biotechnology is the use of biological organisms in technological processes. It is almost as old as the civilization itself, although it wasn't called 'biotechnology' until the 20th century. Far from abandoning it in the 21st century, we are developing new uses for biological organisms. 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
CPEC– A QUICK AND INEXPENSIVE CLONING STRATEGY CPEC is a sequence-independent cloning strategy, making it easy to assemble multiple fragments into any vector and carry out complex library preparations (unlike sequence dependent cloning like Gateway or Univector). It is also flexible as to its use unlike traditional cloning or TA cloning that is restricted in application. SELECTING CELL LINES: ENSURE YOU CHOOSE THE RIGHT ONES FOR One of the most rapidly-dividing cell lines is EB66, which doubles every 12-14 hours. Insect cells like Sf9 and Sf21 double every 24 hours or so, comparable with HeLa (about 23 hours). Don’t be fooled into thinking that all human cell lines grow at a similar rate – HEK293 double every 36 hours, but CACO2 take around 80 hours. THE CS&T REPORT: ITS TROUBLES, AND HOW TO FIX THEM Laser alignment issue. - Clean with 5min clean solution and 5 min of water can help but long monthly clean is better. - Prime. - Check if the short pass filter is in the right position: the numbers should face inwards. - Align the lasers - engineer might be needed. All the values of one laser are off. HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. PAGE 8 – BITESIZE BIO Now that you’ve chosen your perfect primary Schwann cell line, it’s time to culture! Read on to learn how to do so with ease. ReadMore
THE CS&T REPORT: ITS TROUBLES, AND HOW TO FIX THEM Laser alignment issue. - Clean with 5min clean solution and 5 min of water can help but long monthly clean is better. - Prime. - Check if the short pass filter is in the right position: the numbers should face inwards. - Align the lasers - engineer might be needed. All the values of one laser are off. SELECTING CELL LINES: ENSURE YOU CHOOSE THE RIGHT ONES FOR One of the most rapidly-dividing cell lines is EB66, which doubles every 12-14 hours. Insect cells like Sf9 and Sf21 double every 24 hours or so, comparable with HeLa (about 23 hours). Don’t be fooled into thinking that all human cell lines grow at a similar rate – HEK293 double every 36 hours, but CACO2 take around 80 hours.THE FUTURE OF PCR
It’s easy to see potential applications of such systems in areas with poor resources and medical facilities. Lack of electricity, clean water or transportation would not be limiting factors to use this all-in-one technique, with sample preparation, amplification and interpretation of the results done in the same device. This is thefuture of PCR.
ETHIDIUM BROMIDE: THE ALTERNATIVES How can you avoid the perils of exposing DNA to UV light during cloning procedure? Use an alternative DNA stain! Ethidium bromide is not your only option. In this article, we will compare the available DNA stains that can be used in electrophoresis to clarify the options available to you. Ethidium Bromide The classic DNA stain. Ethidium bromide (EtBr) is a flat molecule that fits between METHODS FOR RELATIVE QUANTIFICATION OF QPCR DATA. YES As all of you probably know, methods for calculating relative gene expression from qPCR data include: a) double delta Ct (ΔΔCt) and b) that one other method. Chances are you’ve probably gotten beyond the ΔΔCt method, but you should be prepared in case you face primer sets of different amplification efficiencies. Both methods require the use of a housekeeping gene to control for CHOOSING A COMPETENT E.COLI STRAIN Choosing a Competent E.coli Strain. Learn more. Established in the mid 1970's, New England Biolabs, Inc. (NEB) is the industry leader in the discovery and production of enzymes for molecular biology applications and now offers the largest selection of recombinant and native enzymes for genomic research. NEB continues to expand its product USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most common USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most commonBITESIZE BIO
A Step-by-Step Guide to Designing qPCR Primers. qPCR primer design is a bit of science, a bit of magic, and a little bit of luck. Here’s the science to help you design the best primers for your experiments. By New England Biolabs. DNA / RNA Manipulation and Analysis. INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy A BEGINNER'S GUIDE TO LENTIVIRAL TRANSDUCTION A Beginner’s Guide to Lentiviral Transduction. The use of viral delivery systems to transduce cells for gene and protein investigations has become prominent over the last 20 years. In particular, the use of lentiviral vectors permits stable expression of your gene of interest. This is all possible with a little bit ofnucleic acid magic.
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbedTHE FUTURE OF PCR
It’s easy to see potential applications of such systems in areas with poor resources and medical facilities. Lack of electricity, clean water or transportation would not be limiting factors to use this all-in-one technique, with sample preparation, amplification and interpretation of the results done in the same device. This is thefuture of PCR.
WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
PAGE 8 – BITESIZE BIO Now that you’ve chosen your perfect primary Schwann cell line, it’s time to culture! Read on to learn how to do so with ease. ReadMore
PAGE 9 – BITESIZE BIO Studying DNA–protein interactions is an important aspect of molecular biology. Researches use a variety of methods to study these like the chromatin immunoprecipitation (ChIP) assay, electrophoretic mobility shift assay (EMSA), DNA pull down assay, luciferase reporter assay, filter binding assay, yeast one hybrid system, etc. FIXATION AND FLOW CYTOMETRY Fixation is routinely used in histology and cytology Labs the world over as a way of keeping cells in stasis at a particular point to ensure that, by the time they are examined, they have not deteriorated. This is also something that we often want to do in flow cytometry experiments. It seems like a simple procedure but there are many ways of doing this and, as usual, there isn’t not one THE BEGINNER’S GUIDE TO READING PLASMID MAPS Let’s start with a classic plasmid: pBR322 1. It is often used as a backbone for derivative vectors because it has all features needed for a successful cloning (Figure 1). As you see from the map center, the size of the linearized plasmid is 4361 base pairs. Before you start working with any plasmid, it is advisable to linearize it by cutting USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most common USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTING Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDE Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER) There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most commonBITESIZE BIO
A Step-by-Step Guide to Designing qPCR Primers. qPCR primer design is a bit of science, a bit of magic, and a little bit of luck. Here’s the science to help you design the best primers for your experiments. By New England Biolabs. DNA / RNA Manipulation and Analysis. INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy GOT FFPE? IMMUNOSEQUENCING WITH UNIQUE & CHALLENGING Got FFPE? Immunosequencing with unique & challenging sample types Date: Thursday, June 24th 2021Time: 8.30am Los Angeles, 10.30am Chicago, 11.30am New York, 4.30pm London, 5.30pm Berlin Register Now @ SEQdiscovery In this webinar, you will discover: An overview of the immunoSEQ® Technology and how its quantitative nature makes it uniquely suited to answer questions across A BEGINNER'S GUIDE TO LENTIVIRAL TRANSDUCTION A Beginner’s Guide to Lentiviral Transduction. The use of viral delivery systems to transduce cells for gene and protein investigations has become prominent over the last 20 years. In particular, the use of lentiviral vectors permits stable expression of your gene of interest. This is all possible with a little bit ofnucleic acid magic.
WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
THE FUTURE OF PCR
It’s easy to see potential applications of such systems in areas with poor resources and medical facilities. Lack of electricity, clean water or transportation would not be limiting factors to use this all-in-one technique, with sample preparation, amplification and interpretation of the results done in the same device. This is thefuture of PCR.
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed PAGE 8 – BITESIZE BIO Now that you’ve chosen your perfect primary Schwann cell line, it’s time to culture! Read on to learn how to do so with ease. ReadMore
FIXATION AND FLOW CYTOMETRY Fixation is routinely used in histology and cytology Labs the world over as a way of keeping cells in stasis at a particular point to ensure that, by the time they are examined, they have not deteriorated. This is also something that we often want to do in flow cytometry experiments. It seems like a simple procedure but there are many ways of doing this and, as usual, there isn’t not one THE BEGINNER’S GUIDE TO READING PLASMID MAPS Let’s start with a classic plasmid: pBR322 1. It is often used as a backbone for derivative vectors because it has all features needed for a successful cloning (Figure 1). As you see from the map center, the size of the linearized plasmid is 4361 base pairs. Before you start working with any plasmid, it is advisable to linearize it by cutting USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTINGAGILENT SITE DIRECTED MUTAGENESIS KITPCR SITE DIRECTED MUTAGENESISMUTAGENESIS KIT STRATAGENEMUTAGENESIS PROTOCOL Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDEDIRECT IMMUNOFLUORESCENCEIMMUNOFLUORESCENCE ASSAYIMMUNOFLUORESCENCE ASSAY EXPLANATIONIMMUNOFLUORESCENCE DEFINITIONIMMUNOFLUORESCENCE TECHNIQUEWHAT IS IMMUNOFLUORESCENCE STAINING Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTIONIMMUNO PCR KITIMMUNO PCR PROTOCOLDEFINE PCR ASSAYPCR ASSAY TEST Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER)HOW TO CLEAN BATH FAUCETHOW TO CLEAN BATH MATSHOW TO CLEAN BATH TOWELSHOW TO CLEAN BED SHEETSHOW TO DEEP CLEAN BEDROOMCLEAN MY BED There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most common USEFUL SITE-DIRECTED MUTAGENESIS TIPS FOR TROUBLESHOOTINGAGILENT SITE DIRECTED MUTAGENESIS KITPCR SITE DIRECTED MUTAGENESISMUTAGENESIS KIT STRATAGENEMUTAGENESIS PROTOCOL Let’s discuss some of the comment problems, and the site-directed mutagenesis tips for overcoming them. Getting too many colonies. Not getting any colonies. Getting colonies that have not incorporated the desired mutation successfully. The most important site-directed mutagenesis tip I can offer here is to always, always, always runcontrol
CONTROLS FOR IMMUNOFLUORESCENCE: A BEGINNER’S GUIDEDIRECT IMMUNOFLUORESCENCEIMMUNOFLUORESCENCE ASSAYIMMUNOFLUORESCENCE ASSAY EXPLANATIONIMMUNOFLUORESCENCE DEFINITIONIMMUNOFLUORESCENCE TECHNIQUEWHAT IS IMMUNOFLUORESCENCE STAINING Immunofluorescence staining is a popular and extremely powerful detection method. However, achieving publication quality immunofluorescence or fluorescent antibody staining can get tricky. It’s therefore important to ensure you have the right controls for immunofluorescence. Since immunofluorescence staining is a long process with many steps, it is always advisable to spend some INTRODUCTION TO BIOMARKERS During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more than one definitions. The classic 4 WAYS TO DELIVER CRISPR INTO YOUR CELLS While using expression vectors provides a lot of flexibility, the efficiency is highly dependent on your cells of interest. 2. Viral Vectors for CRISPR. Another option is to introduce the DNA encoding Cas9 and your gRNA into cells using viral vectors. Lentivirus is a popular choice, but other retroviruses (such as MSCV) may be useddepending on
PROS AND CONS OF CRISPR: SIMPLE TIPS FOR WEIGHING UPSEE MORE ONBITESIZEBIO.COM
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTIONIMMUNO PCR KITIMMUNO PCR PROTOCOLDEFINE PCR ASSAYPCR ASSAY TEST Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
SEQUENCING-BY-SYNTHESIS: EXPLAINING THE ILLUMINA The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the originaltechnology.
HOW TO CLEAN A WATERBATH (WHEN YOU CAN'T AVOID IT ANY LONGER)HOW TO CLEAN BATH FAUCETHOW TO CLEAN BATH MATSHOW TO CLEAN BATH TOWELSHOW TO CLEAN BED SHEETSHOW TO DEEP CLEAN BEDROOMCLEAN MY BED There’s something disconcerting about going to incubate a sensitive and irreplaceable sample in a water bath, only to be confronted by the Creature from the Black Lagoon. Unfortunately, water baths are an inviting habitat for all kinds of life, are often shared by many users, and are a perennially unpopular item to clean. To help you overcome your fears of the monsters lurking in the water 10 STUPID LAB SAFETY MISTAKES Keeping safe in the lab really only requires one thing: common sense. But if you look at what people are doing in the lab, you might think that that common sense isn't so common after all. What are the most stupid things you have seen people do in the lab to put the safety of themselves and others at risk? Suzanne and I put our heads together to come up with 10 of the worst and most commonBITESIZE BIO
A Step-by-Step Guide to Designing qPCR Primers. qPCR primer design is a bit of science, a bit of magic, and a little bit of luck. Here’s the science to help you design the best primers for your experiments. By New England Biolabs. DNA / RNA Manipulation and Analysis. INTRAVITAL MICROSCOPY OF CANCER LIVE WebinarIntravital Microscopy of Cancer Tuesday 29th June 2021 LONDON 16:00 BST | BERLIN 17:00 CET | DUBAI 19:00 GST about intravital microscopy techniques that allow you to monitor stem cells and cancer cells in real-time in vivo; how these techniques can help you study the identity and migratory behavior of cancer stem cells; how intravital microscopy uses multiphoton confocal microscopy GOT FFPE? IMMUNOSEQUENCING WITH UNIQUE & CHALLENGING Got FFPE? Immunosequencing with unique & challenging sample types Date: Thursday, June 24th 2021Time: 8.30am Los Angeles, 10.30am Chicago, 11.30am New York, 4.30pm London, 5.30pm Berlin Register Now @ SEQdiscovery In this webinar, you will discover: An overview of the immunoSEQ® Technology and how its quantitative nature makes it uniquely suited to answer questions across A BEGINNER'S GUIDE TO LENTIVIRAL TRANSDUCTION A Beginner’s Guide to Lentiviral Transduction. The use of viral delivery systems to transduce cells for gene and protein investigations has become prominent over the last 20 years. In particular, the use of lentiviral vectors permits stable expression of your gene of interest. This is all possible with a little bit ofnucleic acid magic.
WHAT THE HEK? A BEGINNER’S GUIDE TO HEK293 CELLS There are many advantages of using HEK293 cells. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety ofmethods) and
THE FUTURE OF PCR
It’s easy to see potential applications of such systems in areas with poor resources and medical facilities. Lack of electricity, clean water or transportation would not be limiting factors to use this all-in-one technique, with sample preparation, amplification and interpretation of the results done in the same device. This is thefuture of PCR.
IMMUNO-PCR: A HIGHLY SENSITIVE METHOD OF IMMUNODETECTION Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is adsorbed PAGE 8 – BITESIZE BIO Now that you’ve chosen your perfect primary Schwann cell line, it’s time to culture! Read on to learn how to do so with ease. ReadMore
FIXATION AND FLOW CYTOMETRY Fixation is routinely used in histology and cytology Labs the world over as a way of keeping cells in stasis at a particular point to ensure that, by the time they are examined, they have not deteriorated. This is also something that we often want to do in flow cytometry experiments. It seems like a simple procedure but there are many ways of doing this and, as usual, there isn’t not one THE BEGINNER’S GUIDE TO READING PLASMID MAPS Let’s start with a classic plasmid: pBR322 1. It is often used as a backbone for derivative vectors because it has all features needed for a successful cloning (Figure 1). As you see from the map center, the size of the linearized plasmid is 4361 base pairs. Before you start working with any plasmid, it is advisable to linearize it by cuttingSkip to content
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ONLINE LEARNING RESOURCES FOR SCIENTISTS WORKING FROM HOME From bad weather to rail strikes or global pandemics, there are several reasons you may find yourself working from home. After writing up any outstanding grant proposals or papers, reviewing the literature, and perusing the endless field of coronavirus news updates, what’s a scientist to do? Brush up on your non-lab skills,of course! We…
BY ADRIENNE HUNTRESS Genomics and Epigenetics HOW TO UNDERSTAND CRISPR FORMATS AND THEIR APPLICATIONS Find out how modified variants of CRISPR nucleases provide gene editing with reduced off-target effects and can even control gene expression without altering the DNA sequence. By Sigma-Aldrich® Advanced Genomics Genomics and Epigenetics HOW TO VALIDATE A CRISPR EXPERIMENT Discover how to validate the success of your CRISPR gene editing experiment from determining successful delivery of CRISPR reagents into your cells to the confirmation of desired genetic and phenotypechanges.
By Sigma-Aldrich® Advanced Genomics Organization and Productivity HOW TO PREPARE FOR WORKING FROM HOME AS A RESEARCH SCIENTIST Are you facing working from home? Find out how to prepare and stayproductive.
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Genomics and Epigenetics CRISPR GENE EDITING: CONSIDERATIONS AND GETTING STARTED Discover how to get started with CRISPR gene editing in your experiments with our key considerations. By Sigma-Aldrich® Advanced Genomics Soft Skills and Tools WHAT IS A DOI AND WHY SHOULD YOU CARE? DOIs got you confused? Find out what they are and how to use them.By Laura Grassie
IMAGING AND ANALYZING YOUR WOUND HEALING ASSAY Now that you’ve optimized your setup, you are all set for imaging and analyzing your wound healing assay. ... Making a Mark: How to Set up Your Wound Healing Assay How to Totally Nail Your First _in situ_ Hybridization Controls for Immunofluorescence: A Beginner’s Guide How History Shaped Modern Optical Microscopes, Part One: Simple and Compound MicroscopesTHE LATEST WEBINARS
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CRISPR Gene Editing: Considerations and Getting Started Multiplex CRISPR Gene Editing: What’s It For? Dead Useful: CRISPR-Cas9 Epigenome Editing CRISPR-Based Activation (CRISPRa) of Genes: A How-To Guide CRISPR-Cas9 Genome Editing: Weighing the Pros and ConsARTICLE CATEGORIES
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