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Search below to delve into the Bitesize Bio archive. Here, you’ll find over two decades of the best articles, live events, podcasts, and resources, created by real experts and passionate mentors, to help you improve as a bioscientist. Whether you’re looking to learn something new or dig deep into a topic, you’ll find trustworthy, human-crafted content that’s ready to inspire and guide you.
The resolution of any microscope is related to the numerical aperture of the lens and the wavelength of light used to form the image, and can be calculated using Abbe’s law. This, however, is the ideal situation – the best case scenario. In real life, resolution must be defined in terms of contrast, and there…
Are you stuck in cloning hell?, Tired of doing ligations that don’t work? Want a faster, more efficient cloning procedure? You should try ligation-independent cloning. A growing number of researchers swear by ligation-independent cloning methods because they are simpler and more efficient than conventional cloning and as a recent convert to their ranks, I’d like…
Whether you’re just beginning, or nearing the end of your undergraduate career, getting involved in research can be overwhelming and stressful. There are many factors to consider, so keep it simple and focus on the most important ones: What area of research motivates you? What organisms do you find interesting? Are there any research professors…
Greetings, I’m a research student and I’m having a lot of trouble in finding the right antibodies for my study, because I need both monoclonal and polyclonal. And I need to start addressing some other international scientists. The problem is that I don’t know how to address the situation. If they indeed have antibodies that…
When we are doing a double labelling, it makes good sense to choose two fluorophores with spectra widely spaced apart to avoid mixing. Why combine GFP with Rhodamine, when you can combine it with Texas Red?
Consistent qPCR results depend on careful reagent handling, proper primer storage, accurate pipetting, and contamination control. Running standard curves for each primer pair and following MIQE guidelines ensure reliable, reproducible data. Diluting templates appropriately and making fresh dilutions also improve accuracy. These practical tips help researchers save time and resources while achieving dependable qPCR outcomes.
The most comprehensive way to evaluate DNA concentration and purity is to use both UV spectrophotometeric measurements and agarose gel eletrophoresis. This quick reference guide gives an overview of the information that can be derived from both. UV spectrophotometric measurement of DNA concentration and purity DNA itself, and most of the common contaminants found in…
One of the most crucial steps in any cloning procedure is the preparation of the vector. Get it wrong and your chances of success will be drastically reduced. The overall aim for a good vector preparation is to obtain a fairly concentrated stock of undamaged, fully digested plasmid DNA that is free from contaminants. Missing…
The human brain autofluoresces—a funny thought next time you see a cartoon character with a bright idea and a light bulb over his head—but not so funny if you are attempting immunofluorescence analysis. But there are some significant advantages to using fluorescence detection over chromogenic methods. In this article, I will cover the advantages of…
Ever wonder what those small colonies, like satellites, surrounding a larger E. coli colony on your LB with ampicillin plates were? Or why, when you picked that colony, it never had the plasmid you just transformed? Well, it’s because those satellite colonies are “protected” from the ampicillin by the big colony. Read on for more… Ampicillin…
The concepts of brightness and contrast are so general, and the issues related to them so many, that it may seem strange to have a single brief article with such a title. Indeed, when we speak about brightness, we can think about the brightness of the light source, the aperture and magnification of the lens,…
In the previous installment of this series on western blotting, we addressed potential sources of error when your final product is completely bare. But alternatively, what do you do when too much background is the problem? You may have beautiful bands of interest—but if there is a bunch of non-specific binding, your quantification and data…
Most of us use pretty standard transformation protocols for E.coli. Yours probably goes something like this: – Thaw the competent cells on ice– Add DNA– Electroporate (or incubate then heat shock for chemically competent cells)– Add rich medium (LB or SOC)– Incubate at 37°C (or appropriate temperature) for 30-60 minutes– Spread onto antibiotic plates That…
Dear Aunt Yersinia, A very annoying postdoc in our group keeps telling me off for spinning E.coli at 13K in a tabletop centrifuge. The postdoc claims that high speed damages cytoskeleton and this will reduce my transformation frequency. But I don’t believe her as the cells are cushioned by water during centrifugation. Can you tell…
Problems with expressing your gene? One of the potential stumbling blocks in heterologous gene expression is incompatible codon usage. Every amino acid can be encoded by more than one codon, and for every amino acid each organism has a favorite codon that it tends to use more often than the others. The availability of tRNA…
Discover the different approaches to FRET quantification and get tips for selecting your FRET pairs.
My PhD is rapidly becoming a distant memory. Before nostalgia completely obscures my recollections of this chapter of my career, I thought I’d jot down some pointers for prospective and current PhD students. These are mainly based on things I wish I had done during my PhD, or mistakes I have seen others make. I…
No matter how efficient you are, it’s always possible to improve your productivity and improving your productivity means that you get more of the rewards you are trying to obtain: results, publications… or dare I say it, money. Here are 20 ways to improve your productivity. Some are focussed toward improving the productivity of bench…
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 your two epitopes with primary antibodies from two different species, 2) bind these primary antibodies with two secondary fluorescent antibodies, one…
Dear Aunt Yersinia, I work with a yeast vector where my gene is under a Gal promoter. My boss told me to grow yeast in medium with sucrose, and then add galactose to induce the promoter. I did it, but my protein is not induced; the Western blot is empty. I sequenced a part of…
Ever tried to turn the volume all the way up on a small radio or small stereo system? (Hopefully you have not tried it with earphones in!) Notice how, after some point, the sound didn’t get any louder- it just got more distorted? That’s because you’ve hit the ceiling of your machine’s dynamic range. It’s…
We are pleased to announce that the famous maid of microbiology, dear old Aunt Yersinia, has agreed to start writing a microbiology and molecular biology advice column for Bitesize Bio. She will be free to answer your most pressing questions sent to: auntyersinia@bitesizebio.com By way of introduction, Aunt Yersinia is bestowing 8 spores of knowledge garnered…
“A two photon microscope has higher sensitivity than a normal confocal microscope, because it uses two photos instead of one!” Yes, I can bear witness that this phrase has actually been uttered, and it was not by an undergraduate student. No exception to the rule The condensation of various levels of misunderstandings in this statement…
To answer some of the more interesting research questions, you often need to get a good look at what’s going on inside the cell. Whether you’re running a Western blot or measuring enzyme activity, many assays require access to the materials (e.g. proteins, DNA, subcellular fragments) contained within the cell walls. There are several ways…
A while back, I read an article on Bitesize Bio entitled “When Your Partner is NOT a Scientist” that piqued my interest…for the wrong reasons. And I discovered that I hold polar opposite views on balancing a marriage and an occupation as a scientist. So I was compelled to write this article, not to be…
Whenever you make up a solution or use a purchased reagent in the lab, you trust the work of a whole army of people and equipment. If any one of those links in the chain has a technical problem or makes a mistake, then your reagent might turn out to be faulty. And that can…
RNases are like the baddie super-heroes amongst laboratory enzymes. They are omnipotent, destructive and seemingly indestructible. This is because they were created by evil overlords, for the sole purpose making life difficult for the brave scientists who battle every day to produce high quality, intact RNA preps. Ok, I’m joking about the overlords part. But…
Calculations can be the bane of laboratory work. Fortunately, there are many easy methods to help you do the maths you need in the lab. Here, we tell you about the different ways to calculate primer concentration depending on the starting material. For all calculations, let’s assume we have 22 nmol of a DNA primer…
I have been fortunate enough that in my career to date I have rarely experienced the problem of other people stealing my reagents. However, one PI told me of her experiences working in a US laboratory where things had got so bad people brought their reagents home at the weekends! Working in a research laboratory…
After years of hard work in the lab, having to sit down and write everything up can be a daunting and difficult task. Here are a few tips that I picked up during my thesis write-up that might help to make the process that little bit easier. Back up This is probably the advice most…
Working with RNA? What fun! Those little, nearly indestructible RNases are everywhere – on your skin and mucous membranes, in the water and (some of the) enzymes you use, on lab surfaces, even in airborne microbes! Here are 10 ways to keep the RNases at bay, and keep your precious samples safe: 1. Clean everything;…
In today’s technology-driven world, we leave so many things to our electronic gadgets. Surprisingly, many life scientists try manically to control the appearance of their documents by hand with programs like MS Word. LaTeX takes this task off your hands by providing highly efficient algorithms to properly format your texts. The results are almost always…
I’m sure you’ve heard of Parkinson’s Law, or at least the modern-day generalisation of it. It states that “Work expands so as to fill the time available for its completion.” When I first heard this, back in the mists of time, I thought Mr Parkinson was damn right, had a chuckle at a very accurate…
Site-directed mutagenesis studies can be extremely useful for elucidating the function of a gene or protein, or for creating variants of an enzyme with new and improved functions. There are now many approaches available for generating site-directed mutants, whatever your purpose. In this post, I’ll summarize three techniques that will enable you to produce a…
We scientists are all so focused on getting our work published that many of us seem to forget something very important; that publication is just the beginning. After publication is when our manuscripts really have to do their essential work of communicating our science to our peers. If no-one reads the manuscript, we might as…
Here’s an all-too-often repeated scene in the lab: First thing in the morning, you approach the 37°C incubator with trepidation, open it and through one half-open eye you take a look at the LB plate that you spread your ligation-reaction-transformed E.coli aliquot onto. Looks good – thousands of colonies. Emboldened, you take your “no ligation…
Learning how to speed read can save you a bunch of time; we’ve outlined some simple steps that will dramatically improve your reading speed.
Sepharose beads are porous, which gives them a high surface area for interaction with proteins and allows them to hold a lot of liquid. This is perfect for the application that they were originally designed for: purifying milligrams of protein in columns. When immunoprecipitation (IP) – a small-scale technique for pulling specific proteins out of solution using…
Looking for a new direction in your career? This is the latest in our new series of articles that focus on careers for scientists. When we started this series, we thought you’d be interested in finding out about who does what behind the scenes at Bitesize Bio, and how we got here. I interviewed our…
Looking for a new direction in your career? Well you’ve come to the right place. This is the first in our new series of articles that focus on careers for scientists. We’ll be looking at jobs that are off-the-beaten-track (and sometimes on no track at all) and ones that are certainly not on the standard…
The eBook with top tips from our Researcher community.
