Curse the PCR...

Everything started so nicely… I mean, two weeks ago, everything was on top, my experiments were running, five samples, all positive, subsequent analysis kind of working, not perfect, but working…

Funny thing is, things never worked since and no one has an idea why. I mean, it’s a PCR (for those who don’t know it, polymerase chain reaction, a standard procedure to multiply a specific piece of DNA from basically any sample, blood, cells, feces… ). And it WORKED in the beginning, and seriously, this procedure and also my thesis should be bullet-proof, absolutely, once the PCR works and the analysis works BAM, all easy, all standard procedure.

Well, only that my previously reliably repeatable results are just GONE, and no one knows why. I don’t realize I do anything different than before. It’s the buffers, maybe? Replaced, doesn’t work. The enzymes? Replaced, still no result. The nucleotides? Guess what, I took new ones, no change. My samples? But why THE HELL did they work before? And if it’s the samples, why do they work when a Ph. D. student uses them for the same reaction to test it, but not when I do the EXACT SAME THING? Same for the primers, here we go, checked everything that might go wrong on the material level. Leaves me… Gosh, I feel like the last idiot on earth, unable to reproduce my own results, unable to move on even inches because my very first step does not work. And no one knows why. And everyone tells me the same things I already did. I have no clue. And I feel miserable being unable to proceed on my own, running for help and advice all the time. I can do better than tis, except at the moment it seems I can’t…   I have no idea what’s going on, and it’s been for two weeks now, I still tell everyone everything’s fine and we’ll solve it and I am not freaking out, I try to smile through that all, but hell yeah, this is so frustrating. And it must be my fault but I don’t find how…plus I feel miserable for going onto everyone’s nerves so much because of that sh**…

I mean, it’s not that I don’t like the labwork, people are amazing and the work itself is actually quite fun, it’s okay – it’s in fact the hours of the day I am calmest, concentrating but relaxed, when I am putting small amount of colourless liquids into slightly larger amounts of other colourless liquids and hope that miraculously I’ll see fluorescent bands of DNA of approximately 200 base pairs of length that indicate the product I want to analyse – only it never does at the moment. And the people are just amazing, they are all cool and helpful and it’s a really nice, relaxed but productive atmosphere, so no complaint whatsoever there…

But then, as I said, my stuff isn’t working and no one knows why. For two weeks, And apart from what I tell everyone, it is freaking me out. I felt slightly nauseous this morning, I couldn’t even eat breakfast, which isn’t such a rare thing in the mornings these days, and on top of that I haven’t slept as much as I should in weeks because my (GOD DAMN IT) stupid body manages to wake up beyond any possibility of falling asleep again way too early without need – depending on the day between four and six in the morning, on weekends and in absolute darkness my best shot was eight o’clock after going to bed at one in the night after a day of festival, see what I mean?

Anyway, I didn’t feel like eating and then it hit me like a hammer that I am just scared. I was plain scared to go to the lab – not because I am stressed there, because there is pressure or I don’t like it there, but I was scared to fail again. To not meet my expectations again. To bother all the others again, and again, and again. I am scared to turn on the UV light and find that there is NOTHING. I am just plain scared of failure, and I have no idea what I should do about it. I can’t go running to my professors or m supervisors crying that I am despairing about this and I don’t know what to do. I cannot just run crying to the technicians and Ph.D. students for help, or go whining about how sorry I am to bother them and how stupid and useless I feel. Or that I am better than that by far, usually…

I don’t know… I really have no clue, I guess I’ll just keep trying and waste a whole lot more of material – what fun burning money, for at the moment that’s precisely what I am doing. Never mind that the enzymes alone cost more than I have available in a month… I don’t know. Push on, smile through it and cry on the toilet when no one sees it – suffering from relative sleep deprivation should suffice as an excuse for red-brimmed eyes, I guess…

Man, I am really frustrated at the moment. I think – and hope, and pry – that when I read this in a few weeks’ time I’ll feel angry at myself for letting me so low, especially when it starts working again. I already feel like kicking myself in the butt all the time, which is probably good. Because surrender and giving up and despairing is definitely not an option! This is such a cool project, it just has to work, and it will again! After all, it did two weeks ago… good old times…

3.7.12 21:04

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