#and PI was like /: we try to avoid knockouts bc expensive | Explore Tumblr posts and blogs

raskies456 · 2 months ago

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literally just looking at various potential research labs for grad school and my brain has the audacity to give me the “fucking up the protein purification in front of everyone so badly they all think I’m an idiot” nightmare again

#haven’t had that bad boy since I left my research tech job #to be fair I’m p sure I’m the only person in the labs I’m looking at that has ever even purified a protein #like. of the labs I’m looking at the only wet labs are somewhere that doesn’t have the equipment on site #the others aren’t wet labs at all #but it is funny bc I was talking to a potential PI and she mentioned a problem that I could consider working on and my brain was like. well #this is prob best approached by enzyme activity assays #and I’m p sure this guy can’t be assayed in vivo #so. protein purification for in vitro testing?#but I HATE PURIFICATION.#and also literally if I wanted to do it I’d have to go to the other campus that has the centrifuges and liquid nitrogen!!!!!#which is fucking bizarre to me bc I worked somewhere where those things were standard #the university doesn’t even have an fplc……….#they simply don’t do that shit. which is kinda the point bc I don’t like that shit #and YET #somehow this specific problem offered to me as a potential thesis activates my sleeper agent purification brain #maybe if I were purifying my Own proteins for my Own assay to answer my Own questions……….#like. I wanna study evolution and genes!!!!!!#but if your genes keep having similar deletions in a protein under a certain condition I wanna know why????????#is it loss of function????? and if it’s not how is it changing function???????? how is it benefiting the organism?????#I was like okay well. have you tried knockouts of wild type and subbing in just this mutation #and PI was like /: we try to avoid knockouts bc expensive #and I realized how spoiled I have been bc I was in a lab where we already had and could afford knockouts and double knockouts #like oh you wanna study this gene? wanna sub in a modified version?#cool let’s order primers we have the knockouts you can put it on whatever plasmid expression system you want #that said. we did not have a plate reader or easy access to genotyping esp expression mass spec so #pros and cons. they do different stuff that they are set up for #but also apparently during Covid the prof that studies malaria literally used his own blood bc he couldn’t afford to source it #which. honestly given current events it’s good to know everyone already can buckle their belts and run a lab on two cents and fellowships #456 words #lab tag

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