Technique of the Week
Great tips, tricks or techniques deserve to be recognized. Get the inside scoop on a different protocol each week.
We know the little things matter in science. In fact, we spend much of our time troubleshooting laboratory techniques to find out exactly which little thing is ruining our experiments. It goes without saying that when working with an instrument as sensitive as a mass spectrometer, subtle differences in sample preparation or instrument tuning may dramatically affect the quality of the results. So when things go wrong, we’re not implying you should put every step under a microscope- but here’s what you may find if you do.
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Despite it’s importance, reaction monitoring can be lax, in part because of the hassle of getting into a sealed round-bottom in the middle of a reaction. We have to unclamp, flip-up septum, re-clamp, remove septum, take sample, replace septum, unclamp, seal septum, reclamp then get on with the TLC. After all that work, it’s time for a nap…
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The Video:
The DNA electrophoresis video is in two parts. The first being the overview of electrophoresis and the second being an up close loading of samples in a gel. The reason for the first part is because I needed to fill in time and I wanted to see how well the video would time compress. I like playing around with that stuff, so I just did it. The second part was to take advantage of my new High Definition(HD) camcorder.
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Whether we’ve just started a project or we’re expanding our current one, there are few things more overwhelming than learning about a new gene from square one. Just understanding the logistics can be a killer (Where is it expressed? What other proteins interact with it?…) and usually necessitates reading/skimming/sleeping through a chest-high pile of reprints. Until now…
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Whether brute-forcing through hundreds of mini-preps or setting up so many reactions that you run out of round-bottoms, it’s not unusual to feel like a robot from time to time. It’s normal. However, if that robotic feeling doesn’t seem to be going away and you have a craving for WD40, call your doctor – or mechanic – immediately. Watch this video to see what can happen to an otherwise normal grad student after too much repetition…
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How many protocols can you follow and then actually drink?! Unless you like the taste of silica gel or Tris buffer, probably not many…
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Research is filled with numbers- from activation energies to reaction rates. While many of us can artfully dodge many of the gruesome equations associated with quantum mechanics or enzyme kinetics, one basic calculation remains inescapable.
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All it takes is a little paper cut on your pinky to be reminded of exactly how many times you use the finger that you most certainly have forgotten about. For the next day or two, you will have a renewed respect for the distal digit and realize that in fact it’s a pretty productive member of the hand afterall.
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This protocol is great for three reasons:
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Great techniques don’t have to be expensive or expansive. They’re often clever solutions to routine problems and this Technique of the Week is no exception.
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When I first started cell culture, I was just maintaining a line of HEK cells so it was pretty much meat and potatoes cell splitting. I thought I had it all under control. Then I tried a 1:8 split into a 24-well dish…
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The very first things I ordered when I officially joined my lab were multi-colored Sharpies and a rainbow of lab tape. The color gives me a little bit of joy in a world of black, white, grey, and beige. Yes, I am the kind of person who wants tube racks in every different color. Yes, I am the kind of person who gets excited to use multicolored eppis. Yes, I am the kind of person who enjoys adding NaOH to phenol red solutions just to watch the yellow contrast with the bright pink.
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Weighing out small amounts is always nerve-racking. Between a shaky hand, an unwelcomed breeze or a scale full of static, there are plenty of ways for your precious few milligrams of material to vaporize into thin air. As a result, it is often tempting to get the solid into the “safer” heavier flask where at least it will be protected. However, this is not always the best idea.
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Messing up your own experiments is one thing. Messing up an entire lab’s is a whole nother. One of the easiest ways to make a large group of people furious with you instantly is to contaminate the incubator. It’s the equivalent of lighting their experiments on fire – they should be mad. The bad part is, it’s your fault. The good part is, it’s completely avoidable…
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Find the right box. Crush the dry ice. Assemble the package. Get a FedEx number. Fill out the forms. Schedule a pick-up. Pay for the package. Wonder why there’s not a better way…
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The bump guard lesson is usually learned one of two ways: A) someone teaches you, or B) someone doesn’t. The former is ideal and you may have no idea how lucky you are. The latter looks like this…
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The first biochemistry lab I worked in had a policy that you could not use a miniprep kit until you had mastered the “old school” way. This meant making buffers, phenol-chloroform extractions and no columns – it was an all day affair. (more…)
I’d heard about a mythical method for extracting limonene from orange peel using supercritical carbon dioxide. Perhaps that doesn’t sound that exciting, but the fact that a friend of mine had seen this being done in a plastic tube, in an exhibition hall at a science education conference really got me thinking. How on earth can you get liquid carbon dioxide in a plastic tube? Surely the pressure would cause the tube to explode?
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Ligations can be painful. Let’s just get that out of the way up front. When I was first learning them as a grad student they were significantly more painful because I had no idea how sensitive they were to the amount of DNA added. So as a beginner, I thought “I’m having trouble getting this insert to go in, I’ll just add more insert.” Bad call.
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I sometimes feel like there is only one way to learn anything in lab: the hard way. A lot of little details in lab go unmentioned, yet can make or break an experiment, and you won’t know it until things either don’t work, or go horribly awry. Losing a day and a half because you didn’t realize that all plastic is not created equal falls into the latter category. Losing someone else’s day and a half is, well, infinitely worse.
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