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What do cell lysis, clean dishes, and gallbladders all have in common? Answer: detergents! These useful chemicals can solubilize fats and other proteins in water. They are the key to applications as varied as lysing cell membranes, extracting DNA, and solubilizing proteins for gel electrophoresis. To help you understand these important chemicals, we provide a…
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 is safe, even your pathology samples. Digitizing your pathology samples can help you better organize and manage pathology for later. And…
Whether you’re a grad student or a post-doc, the decision to apply for external funding should be a no-brainer. It is in both your boss’s and your own interests to do this and will undeniably help you in future career pursuits. But before you even sit down to write, it’s important to do your research…
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 known sequences. Think of it as being rather like networking. You know you want to get to know someone so you…
As researchers, we are constantly on the lookout for new and improved ways to analyze, detect and quantify our favorite protein or gene. Luckily, we don’t always need to reinvent the wheel! PCR-ELISA is a good example of where two commonly used techniques have been merged together to create a very powerful analytical tool. What…
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 to the challenges you throw at it. But depending on your protein’s properties and your downstream detection steps, finding the optimal…
Single-molecule localization techniques are another way to achieve super resolution. Discover how the different variants work, and get info on what researchers are doing with them.
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 the numbers you were expecting. In my previous article I touched on a few reasons why your cell numbers might be…
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 organisms can produce interlocking pieces of DNA – so called compatible cohesive ends (CCE). These are pieces of DNA, which fit…
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! So whether you are assessing proliferation by nucleotide incorporation or by dye dilution, here are some handy hints to help you…
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 found and torn to shreds the person responsible for this monstrous act (!!), consider posting the below tips to help those…
Look Ma, No Kit! Unlike with kits, there are no propriety reagents in a DIY protocol. You know what exactly is in each reagent. The DIY approach is catching on!
The literature is bursting at the seams with information about plasmids, sequences, origins of replication and more, and it can be overwhelming to sift through everything (at least for me!) when all you want is to find out a simple fact about your plasmid of choice!! This series of 3 articles aims to take a…
You are finished with your super-arduous experiment. Now the fun part! To “read” your results with a microplate reader, but do you know which microplate reader to use? Or even that there are different types? Have no fear. I am here to help ye, scientific one! Read on for my pointers on picking the right…
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 why. Hardware aborts A hardware abort occurs when an instrument can’t process the information about an event because it is still…
A few years ago, I was doing research in a lab in Ireland. Our lab, among many others, was moving to a new building. Everything was chaotic. Half our equipment was in the old building and the new lab was creepishly empty. The fire alarm went off every few days. There was a constant…
Glucose is the preferred food source for E. coli, however when glucose levels drop, E. coli need to look for other ways to feed themselves. One way in which they accomplish this is to replace glucose metabolism with lactose metabolism. The induction and control of lactose metabolism is complicated and its process has been exploited…
The completion of the Human Genome Project in 2003 ushered in a new era of rapid, affordable, and accurate genome analysis—called Next Generation Sequencing (NGS). NGS builds upon “first generation sequencing” technologies to yield accurate and cost-effective sequencing results. Fred Sanger sequenced the first whole DNA genome, the virus phage ?X174, in 1977. In that…
Antibodies are one of the most important tools in molecular biology. Basic researchers use antibodies to identify, locate, isolate and quantify specific proteins. Clinical researchers use antibodies to target a specific drug (such as a chemotherapeutic agent) to a particular cell. Because of the cell specificity provided by the antibody, a higher amount of antibody…
Trying a new protocol is always a little daunting, especially if no one else in your lab is doing it. So it’s always good to find a tried and tested protocol from a reputable source before getting your hands dirty. Or at least one that can start you off in the right direction. It is…
Now that we’ve learned about the role of apoptosis in good health and disease, it will be useful to know how we can detect apoptosis in cells or organisms. A variety of apoptosis detection kits are commercially available, and here is a roundup of how they work: TUNEL and DNA damage assays The TUNEL assay…
The magnification and viewing of samples using a microscope relies on both the objectives and the eyepieces working harmoniously together. If you buy a ready-to-use microscope, then the objectives and the eyepieces which are fitted as standard will be designed to complement each other. On the other hand, if you are designing and building a…
If I piqued your interest in the first post about my new e-Book ‘The Bitesize Bio Guide to Protein Expression – a Bitesize Bio eBook’ check out this excerpt from the book explaining what an expression system is and how to choose the right one. What is an expression system anyway? There was a time…
An operon is a functioning unit of genomic DNA that contains a group of genes controlled by a single promoter. Put simply, these genes share information needed to create the tools for a particular task so they share a promoter ensuring they’ll all be transcribed together. The lac, or lactose, operon is found in E….
It’s ironic how much folklore and superstition comes with being in science. “That’s a lucky pipette”, “playing Bach for your cells will help them grow”, “always make your own solutions”; we all have our own tips. Some of them might be well-founded others not so much… Tips from trusted colleagues can be very helpful though….
As with some of the greatest discoveries in science, from penicillin to microwave ovens and play-doh, PCR was discovered serendipitously. Thanks to the work of many scientists, including Watson and Crick, Kornberg, Khorana, Klenow, Kleppe (so many K’s…) and Sanger, all the main ingredients for PCR had been described by 1980. Like butter, flour, eggs,…
Cell sorters do not operate by magic, even it looks that way. It’s about the application of physics, electronics, fast computers and formation of droplets. Whether you bring your cells to a flow core to be sorted or you sort them yourself, it important to know how the cell sorter works. Cells can be sorted…
What do these three seemingly disparate studies have in common? After publication, the high-profile findings in each one were questioned due to the presence of batch effects. Batch effects are ever-present and insidious in science, so as researchers we need to always be on guard against them. Keep reading for a run-down of how they…
If efficient cloning is what you are after, you must give Overlap Extension PCR Cloning a go! This restriction enzyme and T4 DNA ligase-free technique is faster, more reliable and easier to troubleshoot than traditional restriction methods. With only two PCR reactions required, you can insert a DNA fragment into a plasmid without spending time…
Super resolution microscopy shattered the lower resolution limit offered by traditional confocal microscopy, enabling us to see more detail. Discover the different super resolution techniques, how they work, and what they are being used for.
It happens to the best of us. You’re minding your own business and suddenly out of nowhere your mastermix is a bubble bath and your primers are enjoying a froth party. Let’s talk about how to deal with these foamy fiends! When you’re making up your mastermix, you could have a variety of ingredients going…
“What Have You Done To My Cells??!!!” This cry of pain from researchers, frequently aimed at core facility operators, is heard after receiving incomprehensible data for an invaluable tube of cells. Equally baffling to the trained user of flow cytometric instrumentation is when data emerges that is either unreliable or inconsistent with the known properties…
A lot of scientists hate presenting posters. Because more often than not, you stand near your carefully designed, full of great results placard, all alone, while there is a crowd gathered two posters down. There are some conclusions you could come to: My own case study In the last two years I presented two posters…
MIQE what’s that? When writing dPCR materials and methods for a paper have you ever pondered what information you should include? This is where the MIQE guidelines will really help. Guidelines for minimum information required for publication of a digital PCR (dPCR) experiment were published by JF Huggett et al. in 2013. These were a…
Ever done a multicolour flow cytometry experiment, run all your controls, done your compensation and then started to analyse your data and realised that you can’t work out where to put your gates? To err is human… When acquiring data on a cytometer, there can be measurement errors due to counting statistics, errors in processing…
Your stomach clenches. Sweat snakes down your torso. The world seems to slow down. You begin the long, terrifying walk down the corridor. Your mind calls out to you to, “Run! Run now!” but you soldier on until you reach the door and knock. There is no escaping the wrath you will evoke when you…
B.I.T.E., or Bunsen Ill-Treatment and Endangerment, happens every day. In the time since you started reading this article, somewhere out there, a Bunsen burner has been mishandled. This is a dark subject and while some flinch at the thought of discussing it (some stories may be too dark for the public to handle) this writer…
I used to love working in the lab on Saturdays. No, I didn’t spend every Saturday in the lab and yes, I did have a life outside of the lab (that’s one reason Saturday work was so great). But there are some great benefits to working in the lab on the weekend: Beat the commute…
Life in the lab is easy-peasy when you are only prepping a handful of tubes. But what if you need to scale up to 10’s or even 100’s of samples? Scaling up your experiments can have some expected and some not-so-expected, leaving you in a lurch, consequences. Read through our tips so you aren’t caught…
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 whether you need to cover your qualifying exam bases, want to avoid looking like an idiot to your coworkers, or need…
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