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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.
You youngsters don’t know how easy you’ve got it. Kits, outsourcing and improved practices are making research easier and easier. At least in theory (who are we kidding?). In the old days things were much tougher, and many wiley old scientists bear the scars, mental and physical, of carrying out techniques that were mind numbing,…
qPCR is a technique used daily in most labs, but the first step, designing your qPCR primers, can be the biggest obstacle to even getting started. Without a good pair of primers, you can’t start asking the real questions and generating data. And sometimes the effort involved in optimizing an assay for high efficiency and sensitivity…
It’s the molecular biologist’s version of ‘I have good news. . and bad news’. The good news is that I amplified the DNA band of interest. The bad news is that I amplified these other bands as well! Oh, and this smear. What to do? Typically you might try and cut out the band of…
Hydrolysis probes, commonly also referred to as TaqmanTM is a very popular chemistry for real-time PCR. In this article I will compare hydrolysis probes with PlexorTM. But first, a quick overview of hydrolysis probes. Hydrolysis probes, an overview Hydrolysis probes are a popular detection chemistry for monitoring sequence-specific amplification in RTPCR. Just like with SYBR…
After Peter’s Introduction to Linux, those of you brave enough to accept the challenge will now have Linux installed on your machine so today I want to highlight to you some of the wonderful free software that is available for biologists using Linux. Before I start, I should mention that virtually all tools of modern…
In my last article I introduced you to the Plexor System. And from that we already know that while in reactions that user SYBR Green for detection, fluorescence increases with accumulation of PCR product, with Plexor the fluoresence decreases. Today I want to compare some other well-known features of SYBR Green chemistry and see how…
A couple of weeks ago, Nick tried to convince us that we should all be using Macs. But why would you want to use a Mac (or a PC) when you could have an operating system that: That operating system is Linux. And I think that it is high time that more bioscientists got to…
In real-time PCR, there are two primary ways to detect amplicons using fluorescent monitoring. One is intercalator-based dyes such as SYBR Green, and the other is probe-based techniques (hydrolysis or hybridization probes). All of these methods share a similar mechanism of measuring increasing fluorescence during amplification. But there is another completely different way to quantitatively…
Most people that do scientific research for a living seem to have mixed feelings about their job. Many that I know are routinely day dreaming of quitting. This contrasts with the well-established, romantic image of the dedicated scientist who loves his/her work above anything else. So what is the real story? Do scientists really like,…
So you finally got your PhD (or your masters or batchelor’s) and you are making the big switch to a small biotech company. You will probably have been hired for the specific skill set that you have built during your training, but now you have to learn to apply those skills to solve real world,…
So you have some blood stored in the -20 °C or -80 °C and you want to isolate RNA from the samples. If you wanted DNA, you would have many products to choose from. But for RNA, your choices are more limited. Obtaining RNA From Frozen Blood is Difficult Why is that? The reason is that most…
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…
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