My PhD is rapidly becoming a distant memory. Before nostalgia completely obscures my recollections of this chapter of my career, I thought I’d jot down some pointers for prospective and current PhD students. These are mainly based on things I wish I had done during my PhD, or mistakes I have seen others make. I hope they’ll make your life easier! If you have any other suggestions, please feel free to chip in with a comment.
This has been doing the rounds all over the web, so I thought I’d post my 10 favorites.
You know you’ve worked in the lab too long when…
The hysteria among molecular biologists about our old friend ethidium bromide has long been an irritation to me. Researchers are rightly wary of this potential carcinogen. More recently this wariness has been whipped up into a witch hunt by companies touting “safer” alternatives and disposal methods. While I don’t for a minute think that we should all throw our gloves away and bathe in the stuff, I think that it’s time for an informed reality check about the dangers, and the myths about ethidium bromide. Read more »
If you do a lot of heterologous protein expression, take a look at Eurogentec’s CherryExpress Kit. Based on the T7 expression system, the CherryExpress vector has a sequence encoding a small red polypeptide (the heme binding part of cytochrome) fused to the promoter. When your favorite gene is cloned in, the resulting fusion protein is bright red, allowing easy identification of clones expressing your gene. Even better, protein expression level can be quantified at 480nm and the highly soluble peptide could even improve the soluble expression of your protein. Tasty.
Of all the of competent E. coli cell strains available, which one should you choose? The choice of strain to use in a given experiment is determined in large part by the nature of the experiment and the set of traits that best fit it. In this article I summarize some of the most important traits and their benefits in downstream applications. Read more »
I love this great time-lapse video entitled “a day in the lab”. If only minipreps were this fast and easy! Read more »
Buying competent cells from commercial suppliers is convenient, provides a guarantee of quality and gives access to strains with a variety of in-built traits that assist with things like maintenance of plasmid integrity (more on these traits later). However, this can be an expensive business. Read more »
Just for fun, here my top 15 reasons for being a scientist. Add your own reasons in the comments below if you so wish.
This is the first in a three part series on the transformation of E.coli. By the end of this you should be an expert on E.coli transformation and on which strains to choose for different applications. If you’re already an expert I hope it’ll be an enjoyable refresher for you. In either case, please comment below if you have anything to add. Read more »
I love shortcuts, and this one is very good. Chun-Ming Liu of Plant Research International has a number of molecular biology protocols on his website, but my favourite is his One Tube PCR Cloning Method. The protocol involves simply putting the vector, insert, restriction enzyme, ligase (and in his case polymerase for polishing the ends) into one tube, incubating at 16°C for 2 days and then transforming. Read more »
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