Recent Articles

pUC18 – Probably the Best High-Copy plasmid in the World!

Today I would like to introduce you to pUC18, a plasmid most noted for its high copy number. In the first article in this series, we talked about how origins of replication (ori) control plasmid […]

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Stop Pushing, Start Enjoying and Get Better Results at the Bench (and in Your Life)

Say Goodbye to Restriction Enzymes and Ligases: An Introduction to Sequence and Ligase Independent Cloning (SLIC)

Top Ten websites to help you with your PCR experiments

Making The Most Out Of Your Commute To The Lab

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Obtain NGS-Grade™ Quality: Extract, Purify, & Fragment Nucleic Acids from FFPE Tissue with the Covaris Process

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Enjoy the full potential of confocal microscopy with ZEISS LSM 880 with Airyscan

Turn Your Pretty Glass Slides into Pretty Digital Slides: Digital pathology cameras from Lumenera

Imaging at the Edge: Improve Multiphoton TIRF Imaging Sensitivity with Titanium:Sapphire Laser Pulse Shaping

Protein Analysis, Detection & Assay

The Nature of Denaturing (Protein Gels, that is!)

You've nurtured your cells for weeks, perfected your experimental conditions, and nailed down all the controls. You've harvested your cells and gently lysed them, now you're ready to look at the proteins. What's one […]

The Five X-Factors in Bacterial Protein Production and Purification

Every protein is unique and thus every protein has its own set of production and purification challenges – many of which cannot be predicted. Therefore to successfully produce and purify your favorite protein you […]

Insane in the Membrane! PVDF vs. Nitrocellulose – Which One Comes Out on Top?

When it comes to Western blotting, there’s no denying it: Your membrane is a key player. After all it is the physical scaffold that holds your precious samples and it needs to be up […]

Antibodies 101: Every Body Needs an Anti-Body!

Antibodies are one of the most important tools in molecular biology. Regardless of what species, cell type, or organ system you are studying, it is almost inevitable that you will use an antibody-based assay […]

Know Your Western Blot Jargon! A Quick Review.

It has been said that "Incomprehensible jargon is the hallmark of a profession" (Kingman Brewster, Jr) and while I cannot speak for other professions, as a biologist I am inclined to believe it. So […]

Microscopy & Imaging

The power of STED microscopy, Part 1: How does it work?

Do you suspect that your favourite protein is doing something really cool? But you cannot see it because your confocal microscope’s resolution is limited. Then Stimulated Emission Depletion (STED) microscopy is what you need! […]

Is There Overlap? Find Out with a Proximity Ligation Assay

Colocalization blues (and reds and greens) Trying to find if and where two epitopes co-localize (or, to be more precise, where they are found in close proximity) may seem easy at first: 1) Bind […]

Enjoy the full potential of confocal microscopy with ZEISS LSM 880 with Airyscan

If you want a 3D fluorescent image you need a confocal microscope. The only question remains, which confocal microscope should you use? ZEISS LSM 880 with Airyscan unlocks the full potential of confocal microscopy. […]

Turn Your Pretty Glass Slides into Pretty Digital Slides: Digital pathology cameras from Lumenera

Is your pathology ready to enter the 21st Century? Do you want to digitize your pathology? Then you need a really good camera, such as those available from Lumenera. Digital pathology is the process […]

Digital Pathology – why you need it and how to choose the best camera for it

Yup, we really are in the digital age…even the pathology is digital. Facebook, Instagram, Tumblr. It has never been easier to take pictures and share them. The digital revolution is upon us and nothing […]

Nucleic Acid Purification and Analysis

Plasmid v Genomic DNA Extraction: The Difference

If you want to isolate plasmid DNA, you crack your cells open and carry out a miniprep, trying very hard not to get any contaminating genomic DNA in your sample. If you want genomic […]

Look Ma, No Kit!: A DIY Method for Isolating Yeast Genomic DNA

I recently moved to a different research institute and was happy to discover that my new lab had not one, but several different kits for yeast genomic DNA isolation. I like trying new kits […]

Better Plasmid Midipreps Part II: What Causes Low Yields?

Recently we received a question from Bitesize Bio reader Sonia after our article How to: Get Better Plasmid Midiprep Yields. She asked: "What could be the problem when one sample gives a good yield while […]

How to: Get Better Plasmid midiprep Yields

I get many people complaining to me about poor DNA yields from commercial plasmid plasmid prep kits. In this article I will explain the main pitfalls in plasmid isolation and how to avoid them. […]

When Glycogen is not Your Friend – Isolating RNA from Glycogen-Rich Tissues

Bitesize Bio has had a lot to say about RNA isolation, mainly because it is one of the most anxiety-producing requirements for molecular biology; especially when you are first starting out (although isolating proteins […]

Next Generation Sequencing

A Beginner’s Guide to Next Generation Sequencing (NGS) Technology

The first whole DNA genome to be sequenced was the virus phage ΦX174, in 1977, by Fred Sanger. In that same year, Sanger developed the future backbone of the “genome era”: DNA sequencing technology. […]

Beware The Bane of Batch Effects

A promising study on using gene expression to develop personalized treatments for ovarian cancer. A report of surprisingly high levels of differential gene expression among different ethnic groups. The announcement of previously unsuspected levels […]

Sequencing the microbiome: re-discovering our invisible allies

Scientific breakthroughs have profound effects on how we view and treat the world that surrounds us. Especially in the microscopic level, scientific discoveries provide the only understanding we have of Nature. One such case […]

Obtain NGS-Grade™ Quality: Extract, Purify, & Fragment Nucleic Acids from FFPE Tissue with the Covaris Process

NGS-Quality DNA and RNA Extraction from FFPE Tissues In this webinar you will learn: about the truXTRAC™ sample preparation system from Covaris how this system produces high-yield NGS-grade nucleic acids Summary: The controlled generation of […]

Lessons on variant hunting in next-gen data sets

Lessons on variant hunting in Next-Gen Data Sets Understanding the general process of how to make sense of next-gen sequencing results as they pertain to human disease. Specifically, when we sequence the whole exomes […]

PCR & Real-time PCR

Top Ten websites to help you with your PCR experiments

Whether you are embarking on a PhD in molecular biology, trying to amplify a new gene, analyzing gene expression by qPCR, or trying to solve a PCR problem, you will probably resort to a […]

Taking the Pain, Challenge and Rage out of PCR: The BitesizeBio Guide to PCR

Polymerase Chain Reaction, better known simply as PCR, has come a long way in the past 30 years. For those of you old enough to remember the not so ‘good old days’ when PCR […]

Mediator Probe PCR

Want a PCR method that gives you high sensitivity and is cost-effective? Then you might want to try Mediator Probe PCR.  This method gives you sensitivity and limit of detection comparable to that of […]

Understand and troubleshoot PCR with The BitesizeBio Guide to PCR

Over the years, I’ve had my fair share of ups and downs in the lab. The latter quite often centering on a failed or plainly weird PCR experiment. As I’ve gone on and become […]

Thermal Asymmetric Interlaced PCR (TAIL-PCR)

What do bunnies, coins and PCR have in common? They all have tails! (ha ha!) Thermal asymmetric interlaced PCR or TAIL-PCR is used to sequence and analyse unknown DNA fragments that are adjacent to […]

Flow Cytometry

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 […]

Using Flow Cytometry for Fluorescence Resonance Energy Transfer

A marriage of sorts Fluorescence resonance energy transfer, or FRET, is often done using a microscope, which means it can be difficult to analyze large numbers of cells in one sitting. One way to […]

Where Are My Cells: Part 2

The golden rule of flow cytometry, especially cell sorting is: ‘Put good cells in and get good cells out’. When you sort you might not get good cells out and you may not get […]

Using Flow Cytometry for Cell Proliferation Assays: Tips for Success

We all know, generally from bitter experience, that experiments don't always work first time and that sometimes the little things that govern success are the things that get left out of that online protocol! […]

Where Are My Cells: Part 1

If there are a million cells of interest in your sample and you pass them through a sorter, you might expect to get a million cells back. But you don’t – and here is […]

Cell / Tissue Culture

Which Cytokine Will I Get? How to Stimulate Human Cytokine-Producing Cells

Cells are like people: depending on their current environment, past experiences and their genetic make-up they will react differently. Treat cells in different ways, and they will produce different cytokines. There are a lot […]

Tips for a Happily Functioning Tissue Culture Room

You walk into your tissue culture room to find a window open, the incubator’s humidity tray empty and a pipette lying on its side in the hood on a used glove… After you have […]

Getting in Deep: How to Deep Clean a Tissue Culture Hood

One of the most exciting aspects of being a biologist is getting opportunities to examine how and why living organisms behave the way they do. We have technology that enables us to obtain images […]

Seven Things That Really Annoy Me About Tissue Culture

The tissue culture facility can be one of the most important places in the lab.  Many researchers spend hours in the hoods isolating primary cell lines and tissue, generating samples for western blot analysis, […]

Mycoplasma: The Hidden Anarchist of Cell Culture

It is the black death of cell culture. Scientists don’t dare utter its name and many a graduate student has fallen victim to its indiscriminate menace. These stealthy anarchists infiltrate quietly but deliberately until […]

Cloning & Expression

Say Goodbye to Restriction Enzymes and Ligases: An Introduction to Sequence and Ligase Independent Cloning (SLIC)

SLIC, or sequence and ligase independent cloning, was developed by Li in 2007 and published in Nature Methods. What makes it a Nature Methods worthy protocol? Unlike other forms of cloning, SLIC does not require restriction enzymes or a ligase! Seriously! Don’t believe […]

Dressing Up Your Oligonucleotide

Oligonucleotides are those smallish bits of DNA or RNA that we rely so heavily on for many of our molecular biology experiments. In their naked form, they are single, inert strands of DNA or […]

Assembling the Puzzle: Cloning with Compatible Cohesive Ends

Consider a jigsaw puzzle. While most of the pieces have a different picture on their surface, all pieces fit together in an interlocking pattern. As unlikely as it may seem, restriction enzymes from different […]

How to identify supercoils, nicks and circles in plasmid preps

One of my favorite things to do with a student the first time they work with plasmid DNA is to have them run an agarose gel containing 2 samples:  uncut plasmid DNA, and plasmid […]

The Basics: How Alkaline Lysis Works

Alkaline lysis was first described by Birnboim and Doly in 1979 (Nucleic Acids Res. 7, 1513-1523) and has, with a few modifications, been the preferred method for plasmid DNA extraction from bacteria ever since. […]

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