Isolating Proteins? Use Histidine and Transition to Metals
To isolate your protein, you are going to need to tag it with something that will allow you to fish it out of the bacterial protein soup. This is where transition metals can help you out!
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To isolate your protein, you are going to need to tag it with something that will allow you to fish it out of the bacterial protein soup. This is where transition metals can help you out!
Scientists today depend heavily on many molecular biology techniques to perform their research. For example, with the advent of next generation sequencing (NGS): scientists are able to look at very minute details, right down to individual genetic sequence variations. However, the increase in experimental complexity means that every extra step becomes more crucial than…
Nowadays, it is no longer absolutely true that applying for a PhD means that you are striving for an academic position in the future. Although a majority of graduate students are certainly still aiming for a career in academia, and will thus benefit from doing a postdoc or two, more and more graduates are pursuing…
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…
Extracting protein from tissue samples and cultured cells is Step #1 in many biochemical and analytical techniques. Before you can do a Polyacrylamide Gel Electrophoresis (PAGE), a Western blot, or mass spectrometry you need to extract your protein. Nowadays, a lot of labs have switched to kits for their protein extraction but these kits can…
As a research intern this summer, part of my project included expressing and purifying a few proteins of interest. Two out of the three proteins posed no problem, but the third caused me to spend an agonizingly long amount of time– setting up new secondary cultures everyday, waiting for them to grow, forgetting to induce…
As a scientist myself, I have always been focused on performing experiments to achieve my sole goal of producing data for a publication. Recently I started paying attention to the waste I solely generate on a regular basis in the lab. Starting from disposable Eppendorf tubes and pipette tips, to labels and Petri plates, it…
Genomic Science has come a long way since the early days of Sanger sequencing in the 1970’s. Today, there are jazzy new sequencing technologies that include fragment analysis, epigenetic sequencing, RNA/transcriptome sequencing and Next Generation Sequencing (NGS). Increasingly these technologies are becoming more accessible, but they still require highly specialized (read: expensive) equipment. Unless your…
The word ‘retrovirus’ evokes images of HIV, the AIDS pandemic and a desperate worldwide effort to defeat this mighty adversary. But on the flip side, scientific research has managed to tame the virus and use it as a tool for advancement. Retroviruses are commonly used to introduce genes into mammalian cells to express or knockdown…
Take a look at the dotplot below, are you happy with the way it’s presented? Do you think that you could recreate that experiment? If you were a reviewer, would you accept that figure? Sure, it’s flow plot, it shows 3 populations of which two are gated. Read many journals and you will see data…
Western blotting uses electrophoresis and antibody-epitope affinity to give a semi-quantitative and (theoretically) clear measure of protein abundance. It’s a long procedure, filled with many steps—and even more room for error. Learning to troubleshoot certain problems is incredibly important for continued success with this technique. So what do you do when your final imaged product…
Flow cytometry is a fluorescence-based technology, as is fluorescence microscopy and confocal microscopy. Fluorescence is fundamental to how a cytometer gathers data, but I am often surprised, as a core manager, at how little new users know about the process of fluorescence. So, this is where I always start the training process. Let’s get physical…
Imagine trying to build a house without power tools: It’s completely doable – after all, people did it that way for centuries – but it’ll take you a lot longer and has limits. Similar to this, modern day biology now has its own set of “power tools.” So while you could do biology the old-fashioned…
We have already looked into the different types of viral expression systems and when you might use one over another in my previous article. But why would you use viral transduction over similar techniques like plasmids? Just a reminder: Transfection is a lab technique where nucleic acids or proteins may be introduced into cells. When…
A cover letter is a letter written to accompany a job application, a grant application, project reports, manuscripts, etc. that explains the purpose of your writing and incorporates certain key points of your application. It plays a decisive role in accomplishing the task it’s been written for, which is why it is so important to…
So it’s time to write up your thesis. By now, you probably consider yourself an expert on your topic or you hate the sight of it (or both!). Either way, hundreds of typewritten pages are all that stands between you and graduation/freedom. One of your first requirements will be to review the current knowledge on…
As a protein biochemist where bacteria were mere workhorses, imagine my surprise when I began work in a bonafide micro lab! I discovered that bacteria could be much fussier than my good ol’ cloning and expression friends E. coli DH5ɑ or BL21. One broth would not do for all, some even required blood! No, no,…
PhDs have been known for their nightmarish effects on students’ psychological wellbeing, to the point that the acronym PhD has also been dubbed ‘Permanent Head Damage’, ‘Philosophically Disturbed’ or ‘Please Help. Desperate’. Doing a PhD is an emotionally exhausting experience rather than being physically challenging. Here are some tips on how to survive the PhD…
As a dual-degree MD/PhD student I spent two years in the medical school doing classroom learning, now I’m in the lab trying to get a PhD, and then in a few years I’ll return for the last two years of medical school and work in the hospital. In this process, I’ve heard a lot about…
Lab work, as we are all aware, comes with many pressures: one of which is productivity. You want to generate as much quality data as possible to meet publication deadlines or perhaps the elusive thesis. Sometimes it may feel like hours spent in the lab don’t match the amount of data produced: for some this…
While it is true that there are some useful websites like SNPedia, or NCBI that can help you find rs codes for genetic variants, sometimes you need that info coming straight from the oven – particularly when you want to look at atypic SNPs or substitutions that have not been validated. So, in this post I…
I’m sure many of us are aware that the world of cancer research is exploding with the idea of cancer stem cells. This exciting hypothesis suggests that there is a small population of cells in the tumor that have stem cell-like properties. These stem-like cells are able to proliferate and differentiate into all the different…
If you are regularly doing ChIP-qPCR, ChIP-RNAseq or luciferase reporter assays to measure protein-DNA interactions, then this article is for you! ChIP experiments can tell you what DNA sequences your protein binds, and luciferase reporter assays predict whether your protein functionally binds a specific promoter to activate transcription – but a yeast one-hybrid (Y1H) assay…
When it comes to registering the signal output of your Southern/Northern/Western/probe hybridization, you are spoilt for choice these days. You can go all retro and use X-ray film. You can go digital and use a phosphorimager. Finally, you can go fluorescent and use a fluorescence detector. So, what are the pros and cons of each…
Having just finished graduate school, I have been given the privilege of nearly unlimited time to reflect (Yay! Unpaid, Boo!). Graduate school was, for me, a juxtaposition of intellectual growth, real-world learning and great fun. An introduction to adulthood with training wheels—while simultaneously being a blur of anxiety, work, sleepless nights and existential crises. I…
Choosing a PhD topic can be very hard. There are a lot of things to consider from the subject to the supervisor. Here are some tips to help you choose. Find out what you really like This is the first topic because it is the most important. My first advice would be to get some…
If you find it very difficult to call your boss by her first name, feel obligated to wear business casual clothes to the lab, or reckon that it’s slightly weird that your fellow PhD candidates are inviting you to a casual night out instead of to the library to study, then like me, you just…
In Part 1 of this article, I introduced you to using code for basic image manipulation in ImageJ and working with the command recorder to expand your coding vocabulary. I covered how to make a simple macro, how to edit it and then save it to be run again another time. If you skipped the…
In the frenzy of today’s scientific publication landscape, it is essential to take a proactive stand at your online visibility and reputation. There are online tools that can help you make yourself spotted and translate your hard work and high-quality science into actual impact. So if you are interested in improving your chance of landing…
Pat yourself on the back, you saw a post with the word “coding” in the title and you didn’t freak out or glaze over. That’s the first step. Coding seems to have such a stigma attached to it; people tend to think that it’s incomprehensible nonsense that they could never learn, and that it has…
Working in a lab can be stressful, there is so much to do, and so little time to do it that we often feel overwhelmed. Sometimes you come into the lab and there are 15 experiments that just need to be done, so you throw on some music and start doing a bunch of them…
Basic Local Alignment Search Tool (BLAST) remains the cornerstone of biological sequence analysis. When users have one or few queries, the data are easy to interpret based on the graphical output that the search provides. However, when you have to analyze hundreds or a few thousand queries, running graphical outputs can be computationally intensive, difficult…
When you first start out using a microscope, you might only adjust the eye pieces, objectives, and the focus controls. However, you shouldn’t overlook the microscope condensers as they are an important part of the whole optical system of a microscope.
Read our top tips on how to write a scientific review, from gathering your information to getting it published.
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A disruptive sequencing technology Every new generation, a new concept is born and can completely reshape the landscape of biomedical research. Nanopore sequencing technology, although still at its infancy, is beginning to look like a “game-changer.” It’s a revolutionary concept in sequencing in which strands of nucleic acids are fed through a tiny pore (nanopore)…
Want to save money on one of the most expensive steps of your Western blots? Then, use less antibody for significant cost savings!
You open the incubator in the morning and to your dismay there are a hundred glorious colonies… on your vector-only control plate. While there are a number of potential causes, I’ll highlight a few of the more likely culprits and their solutions.
You had been happily measuring your DNA samples with your NanoDrop for a while, until… you noticed that your results are not reproducible *gulp*.
Read on – here are 10 points that will help you troubleshoot the issue.
If you want to know what is going on in the brain of drosophila you can use neurobiology imaging techniques to get a global whole-brain perspective. However, such techniques are slow compared to the rapid nature of the neuronal electrical activity, which may be better studied using Drosophila electrophysiology. Lab techniques are often shrouded in…
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