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Here’s a quick guide to cleaning and maintenance, to help you overcome your fears of the monsters lurking in your water bath.
Epigenetics is the study of heritable changes in the phenotype of a cell or an organism that are not encoded by the genome (hence epi which means ‘above’ in Greek, and genetikos which means ‘origin’). In this article, we’ll discuss DNA methylation, a common epigenetic modification: what it is, how to detect it, and how…
Everybody is different when it comes to how they like to run their work space. Some people just find a space in the lab and work wherever there’s room, sharing their tips and pipettes, while others like to have a designated area to do their lab work in, with their own equipment. Personally, I much…
There are six little words that can instill both excitement and trepidation in the heart of a graduate student: “No one’s ever done this before.” What those words really mean, of course, is “No one’s ever published this before,” and you are either standing at the edge of a great discovery or a chasm of…
When running a quantitative Western blot, it’s crucial that your sample preparation is consistent. Incomplete protein extraction from one sample will skew your results when you compare it to the protein content of a sample that was extracted more thoroughly. And after the protein extraction, it’s important to handle the samples in an identical manner…
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:
When it comes to choosing a molecular weight marker to run on your SDS-PGE gels, there are a lot of options out there. How do you know which one is right for you? Read on for tips on what to consider when choosing a standard for your protein gels. Before you go about selecting a…
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…
So, you’ve done your experiment, prepped your samples, and run your SDS-PAGE gel. Now it’s time for the all-important transfer step, that tricky point that will determine the quality of your Western blot. Transfer times are empirical and based on your own particular samples, which means that there is no easy way to determine how…
The easiest way to keep abreast of what’s going on in your scientific field is to set up a PubMed alert. We show you how.
I think that transferring Western blots is one the most enjoyable tasks to do in a lab: it’s quick, it’s messy, and on some gleeful level, it feels like a child’s art project gone wrong. Of course, it’s also finicky and slippery and prone to tiny pitfalls that can noticeably affect the quality of your…
For Western blot data to be reliable, it is important that you load known amounts of sample into each lane of the gel. This is of particular importance if you are doing a quantitative blot, where you really need to be able to compare band intensity in each sample. In this article, we’ll talk about…
You have just finished your undergraduate studies, or perhaps you have been working in the field for a while, you are full of energy and zest and you want to prove to the world that you are carved from the right stone to pursue PhD studies. Going to graduate school is a very important, potentially…
While Luria-Bertani broth (LB) has long been the fuel that powered Molecular Biology and Biochemistry, there is an increasing movement towards more specialized and complex bacterial media formulations such as Terrific Broth (TB), Plasmid DNA Media (PDMR), and Autoinduction Media (ZYP-5052). These media formulations optimize E. coli cell growth and performance utilizing specialized carbon sources…
I recently introduced you to the concept of polarising microscopy. Naturally, if evaluating refractile material is an everyday part of your research, it is definitely worth investing in a professional polariser modification for your microscope. But if you only use a polariser occasionally, this might not be the best use of your lab’s money. In…
Tandem affinity purification is a development of existing techniques for purifying protein complexes from cells in physiological conditions. It was first described over ten years ago and has become a commonplace laboratory tool. In this brief article I’ll introduce the basic technique and describe some of its advantages. Biology is a team game. Most biological…
When you stop to think about it, tissue slices for immunohistochemistry (IHC) undergo quite a lot of handling. From chemical reactions to washes – even manipulations and transfers between baskets and microtubes – final analysis is often hours away from the initial step of taking a tissue slice. Properly fixed tissue has to be robust…
Polarising microscopy involves the use of polarised light to investigate the optical properties of various specimens. Although originally used predominantly in the field of geology, it has recently become more widely used in medical and biological research fields too. Polarising light microscopy is a contrast-enhancing technique to allow you to evaluate the composition and three-dimensional…
Against the advice of journals and printers, many scientists use Microsoft Powerpoint to assemble posters and figures. You should consider upgrading to Adobe Illustrator! For generating scientific figures, Illustrator is more powerful and flexible than Powerpoint and is designed to produce print documents at high quality resolution. This means that journals will stop sending your…
There’s more to mammalian cell culture than just making sure that your cells don’t die. It is a lot like taking care of children. You have to feed them, make sure that they’re growing well, and keep them under constant supervision. If the cells are put through extreme conditions (over-confluency, media-deprivation, inaccurate temps, etc.), their…
Do you use human cell lines in your research? Well, keep reading because this may be the most important article you will ever read in your research career. It is estimated that 18-36% of all actively growing cell line cultures are misidentified and/or cross-contaminated with another cell line (1). For researchers, this could mean that…
Collecting the data took several years, writing the paper took several months, assembling the figures took several weeks, and converting those figures to PDFs took a frustratingly long day. You waited a month for the paper to come back from review, then two months re-doing experiments to satisfy a sadistic reviewer. Finally, your paper is…
To pull together our discussions so far on hypothesis testing and p-values, we will use the t distribution as an example to see how it all works. The t distribution (you may have heard it called Student’s t) is a probability distribution that looks like a bell-shaped curve (or normal distribution). If we sample repeatedly from…
Now we come to the third part of our trifecta; in the last two posts I have gone over p-values and how they determine significance in null hypothesis testing, and we talked about degrees of freedom and their effect on the p-value. Finally, we come to pseudoreplication: where it can all go terribly wrong. Replication…
In the previous article in this series, we covered teamwork and networking. Now it’s time to move on to what many people consider the most boring part of the lab work: the analysis. I know we all wish that a simple histogram or a rather nice-looking Western blot or PCR would suffice. But the fact…
In the last post I talked about p-values and how we define significance in null hypothesis testing. P-values are inherently linked to degrees of freedom; a lack of knowledge about degrees of freedom invariably leads to poor experimental design, mistaken statistical tests and awkward questions from peer reviewers or conference attendees. Even if you think…
In previous articles, I’ve primed you on hypothesis testing and how we are forced to choose between minimising either Type I or Type II errors. In the world of the null hypothesis fetish, the p-value (p) is the most revered number. It may also be the least understood. The p-value is the probability, assuming the…
Microarrays are one of the most in-depth ways of determining cellular gene expression levels of thousands of genes simultaneously. They are able to help determine: Getting good microarray results starts way before analyzing your data, however. A crucial first step in setting up a good microarray experiment is extracting high-quality RNA from your cells. In…
For applications such as site-directed mutagenesis, it is often recommended that you use a proofreading polymerase (also known as high-fidelity polymerases) to minimize the risk of introducing unintended point mutations. But what is a proofreading polymerase? What makes them different from other polymerases? And when should you use them? Read on to learn more… What…
Before starting any new research project, it’s essential that you have as complete an understanding as possible of the current research literature. Knowing what other people have done will prevent you from duplicating existing work, and will perhaps indicate under-explored niches. If you work in the same subject area over a number of years, you…
Most site-directed mutagenesis protocols strongly recommend that you use only PAGE- or HPLC-purified primers to mutate plasmid templates. Using purified primers is supposed to minimize the introduction of unintended mutations, thus drastically improving the probability of generating your desired mutant. However, specially purified primers can be extremely expensive, and take longer to synthesize than standard…
RNA interference (RNAi) may have originated as a defense mechanism to protect cells against foreign genes introduced by viruses. This concept has since been put to use to create a powerful experimental tool for investigating gene function in organisms. Small-interfering RNA (siRNA) libraries for investigating genome-wide function can be produced by chemical synthesis of probes…
ELISAs (enzyme-linked immunosorbent assays) are often used for detecting and quantifying substances such as peptides, proteins, antibodies and hormones in research and diagnostics. Today, a wide range of ELISA formats exist to suit your needs e.g. indirect ELISA, direct ELISA, competitive ELISA and sandwich ELISA. While it has become easier to perform ELISAs, thanks to…
You have been toiling away at your thesis project for years and you think the end is in sight. Now the big question is “What’s next?” If you think you might want to move away from the bench, then you should check out our suggestions for alternative careers for scientists. If you think your future…
Merriam-Webster defines networking as “the cultivation of productive relationships for employment or business”. Less formally, networking is actively communicating with the other people you know (mostly scientists, in our case) for career advice and job openings, in addition to utilizing opportunities to meet new people for the same purpose. This is a core activity of…
Although the microscope is probably the most commonly used biological instrument, it is frequently used improperly. The rate-limiting step to getting high quality microscopic images is illumination of your specimen. When you examine a specimen under the microscope, the intensity and distribution of light must be clear and equal to enable you to evaluate all…
Several years ago as a freshman in a research lab, the very first project I received was to pipette incremental micro-volumes of H2O onto a piece of parafilm. Boring! Weighing the liquid on parafilm and comparing the weight between 10 replicates for each micro-volume continued for a week before I touched anything else in that…
You might be proud of your pipetting skills (if not, check this article on how to stop pipetting errors from ruining your experiments) and be churning out data faster than a liquid handling robot, but beware… you might also be pipetting yourself out of a job. I almost did. Pain due to pipetting is common….
The presence of supercoiled plasmid DNA on a gel can be inconvenient for molecular biologists, especially beginners because it is easy to misinterpret. Read our top tips how to recognize supercoiled DNA and keep it from derailing your cloning experiments.
If you’re starting your PhD or post-doctoral work, chances are you’ll need to use a light microscope at some stage during your research. Some of you may be seasoned microscopists. For many of you though, this might be the first time you’ve ever plugged in a microscope, or at least the first time you’ve used…
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