i arrived at lab with such purpose. i was going to purify rna, quantify it, and make cDNA out of it, all before leaving for class at 2:00. furthermore, i was going to do it all without assistance. since i was there bright and early (well – it was 9:00), i was in no mood to sit on hold until my grad-student-mentor-guy arrived at some unpredictable hour. i was ready for action, and i figured i could consult manuals and other grad students if i got stuck.
i felt ready to be independent. i felt good.
the rna purification took until about 12:00, a bit longer than i thought. so many spins and washes! but i think i managed to get through it without mixing up tubes or accidentally spilling my final eluted samples down the drain.
i was a little hungry, but still feeling good at this point. gsmg had arrived, but i didn’t feel the need for a consultation. i knew what to do, right? i made dilutions of each sample (there were 20, damn it) to use for quantification. quantification sounds fancy, but it’s not. essentially, all you do is squirt your diluted samples into a little clear holder and let a spec machine do all the work for you. it essentially ‘reads’ how much rna is present based on absorbance of different wavelengths of light.
i was in a very precise, careful mood, so i decided i would read each sample not once, but twice. the machine, though very smart, is not super-efficient. it is designed to read only one sample at a time, and the read is not instant. so forty times i loaded the machine, pressed a button, waited for it to whir and churn out a number, and sucked out the sample to start again. it took almost 2 hours.
the values were looking all right. not perfectly equal amounts of rna, but sort of within a normal range. a few samples seemed to be deficient – i figured probably an error in making the dilutions. so i printed everything out and was about to show it to my gsmg (honestly, i was going to brag about how high-yield my purification had been) and i realized:
the absorbances were totally off. and the concentrations i had generated were not the usual 100 to 200 ug/ml range but were negative numbers. i had forgotten to zero the machine.
so yeah, i wasted 2 hours. i now have to start again tomorrow, loading and reloading the stupid spec machine. by the time i realized in horror what i had done, it was time to leave for class, so i accomplished neither the quantification nor cDNA reaction that i set out to finish. at least i made some rna. and i suppose it was a learning experience:
1. it’s ok to have people remind you how to do things if you’re not completely sure
2. on the other hand, it’s actually not the end of the world if you do something really stupid
3. don’t ever forget to zero the spec machine
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i am thinking about getting a second job as a kaplan mcat instructor. apparently, you can work as few hours as you want, which would be ideal, since in my case, i think i’d want a maximum of 4/week. i think it would be a good experience in learning to teach, too, but with much better pay than a ta position. i know i’m not always going to have as much free time as i have now, but if people can have babies and raise them while completing their phDs, i should be able to handle an mcat class or two. but i’m getting ahead of myself. i have to convince them to hire me first!
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i feel bad not mentioning the last debate. i did watch (though i sort of fell asleep at the end somehow); i just don’t have much to say. just like the other debates, i wanted to repeatedly throw things at the tv screen while bush spoke. both candidates seemed to have their claws out. the polls are saying kerry won the debate, but it no longer matters. the election is what matters. i can’t wait for it to be over.
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