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The following question was emailed to Bitesize Bio by Beheroze Sattha and I gladly took up the challenge, and I immediately knew the answer. Or so I thought. After delving extensively into Pubmed, Genes V (I know, I need a new version) and Molecular Cloning I have come up with an answer, but it is…
If you ask any finished graduate student, most of us starting a Masters or PhD program were very excited at what awaited us and if you were anything like me, you were foolishly idealistic and thought you were going to pull on a lab coat, cure cancer and save the world. Ok, maybe most people…
Criticism is not just valuable, it is essential for a person’s development as a scientist, or anything else for that matter. Well that’s not entirely true. Not all criticism is valuable, it has to be the right kind of criticism. It has to be constructive and better still, well delivered in order to inspire the…
Thanks to Bitesize Bio reader, Muthu Arumugam for contacting us about some problems he has been having with restriction digestion and clean up of DNA. I have boiled his query down to four main questions that are pertinent for most molecular biologists, so I hope that Muthu and everyone else can learn something from my…
In response to my last article, The Taq behind PCR, one of our readers, Bonnie Barrilleaux, asked whether DNA could naturally survive at temperatures that would denature it. It also begged the question; how do proteins stay intact and functioning at these high (55°C and up) temperatures? It turns out, cells do a lot of…
It seems there is a reality TV show for virtually every type of person or profession. From Alaska king crab fishermen to surviving the outdoors to living the life of a privileged housewife, you name it and there is a show about it. So why not a show on the challenges and antics of people…
Rushing and overloading yourself in science is common, even normal. Surprisingly it is considered as an admirable aspect of “scientific flair” in some quarters. And the sad fact is that it is an ingrained part of the scientific landscape. But is this the best way to do things? Should you allow yourself to succumb and…
Gel extraction — what could be easier? Now we have quick and easy gel extraction kits, we no longer need to use time-consuming old fashioned methods like electro-elution or “freeze and squeeze”. Thank goodness. When Gel Extraction Goes Wrong But even the simplest of procedures can go wrong. Maybe you were distracted, confused, or thought…
Controls are obviously extremely important when setting up experiments. Without them, a meaningful interpretation of the experimental results would be impossible. I say obviously, but in my previous job as a technical services scientist, you’d be surprised at how often I found myself talking to customers about the importance of controls. One customer commented, during…
The reverse transcription (RT) step of RT-PCR for converting RNA to cDNA is critical for accuracy in quantification and for finding low copy messages. Thus, you want to make sure that this step is performed with the highest efficiency but without having to optimize every single step. To help you further in optimizing the RT…
Really great presentation skills. Some people in science seem to have them, and some don’t. I am one of the don’ts. Sure, I can get up in front of people and talk when needed, but it won’t be a polished performance by any means. I can get my message across but my delivery is not…
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 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…
Nobel Laureate Kary Mullis is generally credited with inventing the polymerase chain reaction, but his discovery owes a lot to a microbiologist who loved to travel, some refuted assumptions of what can live in hot springs, and a now-closed field station in Yellowstone National Park.Here’s the story. In the 1960s, Thomas Brock was a biologist…
Removing unwanted DNA from vectors is one of the most routine laboratory techniques in cloning. Check out our top tips.
No matter how many times you look at it, it’s not going to change. You are planning your next cloning experiment, but there’s a problem. The only restriction enzyme that cuts in a suitable position on your plasmid vector also, as luck would have it, cuts in another position elsewhere in the vector so you…
As well as having had some negative work experiences, I’ve also had the pleasure of working with some wonderful people, including some of my previous bosses. Life is too short to deal with some of the idiosyncrasies described in Suzanne’s previous article on bad bosses.So let’s balance the scale and look at what it takes…
Now that you know who you are trying to impress with your resume, it’s time to focus on how it looks and reads. Remember,.this is NOT the venue to display your creativity. They key is to keep it simple and clear. Follow a simple format that is easy to read – remember that your audience…
Going to conferences normally involves a significant investment of time and money. So it’s important to get as much as you can out of them. One of the most valuable things you can get from a conference is contacts. These can build into a network of people that will be valuable to you throughout your…
RPM and RCF are two units that can be used to describe the speed of a centrifuge. Although they may look similar, they are oh-so-different and confusing them has resulted a disastrous end to many an experiment. So let’s set it out in black and white to make sure you don’t succumb to the same…
In my last article, I introduced Cell-Free Protein Synthesis. Today I want to talk about a major bottleneck in in vitro cell-free protein expression; low yield. Most often, paying attention to the important factors such as template purity and design, transcription and translation inhibitors, and potassium and magnesium concentration will solve any problems with low…
After ligation, the method you use for desalting your sample prior to electroporation is critical, especially if your ligation is inefficient, according to a study by Schlaak et al [1]. Under standard electroporation conditions, the electric field of 12-18 kV/cm generated in a 0.1mm-gap electroporation cuvette means that the conductivity of the sample must be…
If you search the literature using a comprehensive search engine like Google Scholar, you will get several types of articles listed. Most of them are peer reviewed journal articles and many are patents. But beware of an important distinction between the two: Although patents can contain useful information, they are not authoritative because they are not…
Looking for cheaper or faster solutions in the lab? Here’s our top 10 list of ways to use everyday items to make gadgets and for low-tech solutions for the lab.
Undergrad courses teach you to learn in a specific way. You have to cram in as much information into your brain as possible, hold it in there, then regurgitate as much of it as possible on exam day. Of course, actually understanding what you are talking about, and working from basic principles, helps but the…
Cell-free protein synthesis (aka In vitro translation) refers to protein production in vitro using lysates generated that provide the cellular machinery necessary for synthesis. The lysates can be of bacterial or eukaryotic origin. It is a useful alternative to in vivo synthesis for generating protein for the study of things like: So what are the…
There is nothing more frustrating than getting back rubbish data from a DNA sequencing run, especially when you are waiting for an important result. For example, confirmation of that clone you have been trying to get for the past three months! A lot of the time, the quality of sequencing data is within your control….
qRT-PCR (quantitative reverse transcription-polymerase chain reaction) is now the gold standard technique for mRNA detection and quantification, sensitive enough to enable quantification of RNA from a single cell. The reverse transcription (RT) step is the main source of variability in a qRT-PCR experiment, so an optimal reverse transcription is essential for a reliable and successful…
No scientist is an island, not even a great scientist like you. A good network of professional contacts is as essential to your career as hard work at the bench. This is because your network can open many doors that no amount of good bench work could. Access to unpublished information, collaborations and job opportunities…
Transfection of eukaryotic cells is a routine but sometimes tricky procedure. There are several transfection reagents available on the market, but sometimes the old methods are the best. I find that the simplest, fastest and cheapest transfection method for eukaryotic cells is calcium phosphate mediated transfection (1). It’s main advantage is that, since Ca2+ is…
Heat maps are a useful way to represent certain types of data; the data are colored by coloring according to the values in them, (e.g. red for high values, yellow for medium and green for low values), providing a powerful visual representation of a data set. This allows you to quickly see results from DNA…
PapersIn recognition of a fresh new year and a fresh new direction for Bitesize Bio, I thought I’d start my tenure as the fresh new Bioinformatics “guru” by introducing you to a handful of websites that will help you locate established as well as fresh new and (mostly) free online molecular biology and bioinformatics databases…
Everyone is worried about getting results, aren’t they? Results are what you need for success in science – they are essential for bringing the funding in. But focusing on results per se is not a good way to work because, as a scientist, you can’t “get” results. You can’t “make” them happen. Essentially in every…
In my current position, I had the opportunity to hire a new R&D scientist to join the team. I was excited at being able to build my team and take my time to find the perfect fit for our company. My experience in the process of hiring gave me a new perspective on all the…
You are not alone. Everyone makes a hash of their protein gel sometimes but this resource can help you work out what went wrong, and feel better for seeing gels even worse than yours.
Your DNA samples are precious so take care of them! Here are 5 ways that DNA can be damaged, so now you now what to avoid in the future.
Ever heard of polymerising agarose gels? I haven’t. If you think you have, read this.
DEPC. IF you work with RNA, you’ll know this stuff. It’s vital for ridding solutions of RNases that would otherwise destroy your work. But just how well do you know it?
There’s more than one way to do a plasmid miniprep. Here are 5 to add to your molecular biology arsenal.
This is the story of how one of the most famous and quirky naming conventions in biology came into being.
Calculating your fudge ratio can help you get your work done on time and get home before dark. Here’s how.
The eBook with top tips from our Researcher community.