A Week in the Life of a Molecular Plant Pathologist . . . or the adventures of “Gel Woman”
Everyone warned me that a Ph.D. in molecular biology would involve countlessfrustrations, and that I should not expect to get a “Result” until atleast the final year. Undeterred, I began the task two-and-a-half years ago,determined to pursue the challenge to the bitter (sweet) end. Not only did Ichoose molecular biology as the trendy but fiddly tool for my investigations,but the object of the study was to be the enigmatic, obligate biotrophic fungalpathogen of wheat; yellow rust. A sucker for punishment, or what?
Typically, the week starts off at a moderate pace, tying up the ends of lastweek’s experiments and deciding where the hell to go from here. Apart fromalways being busy, and the seemingly endless PCRs, there is no such thing as a “typical”week. Each is unique in the variety of successes, failures, and complicationswhich spring up. Planning experiments usually starts after coffee on Monday,after I have developed an AFLP auto-rad which invariably would be better suitedfor sunglasses than thesis fodder. Coffee time (11 a.m.) in the Plant ScienceSection is important to catch up with the goings-on over the weekend, but I findit best to avoid the inevitable football conversation by checking that there isnothing worth watching on telly tonight.
Back in the lab, I make up some media to autoclave, and attempt to solve theproblems of an exam-frenzied undergraduate who picks this moment to come lookingfor Matt. Matt is my supervisor, who has that extraordinary power possessed byall PhD supervisors by which he knows exactly when his help is needed so that hecan vanish, only to reappear once the panic is over. Later, after alunchtimePostgrad Committee meeting (I have the dubious honour of being PlantScience Rep), I pour some plates, and then go down to the glasshouses to treatmy wheat seedlings, collect spores (not always forthcoming), and attempt tocheer up the glasshouse staff (likewise). Then I run a gel.
Last week I started early on Tuesday so as to plate out my cDNA library forsecondary library screening. I was just about to take my bugs down to theculture room when Cristiana, an Italian student of notable dress sense, bouncedinto the lab wearing her Superwoman top and announced that, after her holidayand exams, she was now ready to repeat her PCR. I showed her where to find theTaq, and how to programme the Robocycler before I started to set up mycultures. After lunch I sat in the lab listening to the radio, purposefullypipetting PCR samples, and checking the incubator occasionally to see if therewere plaques appearing on my library plates. This happened eventually at 7.30p.m.
Wednesday is usually Seminar Day, but last week it was Postgrad SymposiumDay, and this meant me. I was strangely calm in the morning and I ran agel with last night’s PCR reactions. There were bands in the lanes I had hopedfor, but I will need to test more isolates. Then I made plaque lifts from thecDNA library plates – one of my favourite tasks, especially in the summerbecause it’s a good excuse to work in the cold room. I ate my sandwiches, readthrough my notes for the umpteenth time, and began to get nervous. The symposiumstarted at 2 p.m. and despite the chairman’s failed attempt to pronounce “Pucciniastriiformis“, I started off confi dently: once I had described thelife-cycle, pathotype nomenclature and control methods I was over the trickybit, and it was plain sailing into the molecular stuff and my “Results”.Afterwards, everyone seemed quite impressed and commented that I’ve actually gotlots of “Results” already. Hmm, but are they enough?
The pace of activity increases on Thursday, when I fret that there are notenough days left this week. I do a plasmid prep on the cells which I resurrectedfrom the ‘fridge the previous day, to see whether I managed to clone anythinglast week. I will run them on a gel after I’ve done a restriction digest. IfCristiana is “Superwoman”, then I must be “Gel Woman”.Thursday is the day the radioisotope comes, so today it’s probe- making for thelibrary. Matt knows how much I hate making radioactive probes, so he comes tolend moral and practical support. After lunch I can set up the hybridisation,but first it’s time for a crazy game of badminton (a quick change into myfreebie AFLP tee-shirt and I am Gel Woman!) – the aim: to totallyde-stress.
Friday morning is fraught with activity. I wash the filters and monitor thembefore I put the film down. They are quite “hot”, so could this be the”Result” I need? Matt takes this opportunity to come into the lab andshow me a paper that he has found about a PCR test which may be useful. It’s allreally exciting but I am too busy to be excited today: there is washing up to bedone, plants to be inoculated, my lab book to write up, things to order and gelsto run. Finally, before I leave for the weekend, I write a list to remind myselfwhat will need sorting out on Monday morning. Now, down to the pub . . .
Katherine A Steele
University of Nottingham