April 28th, 2007

meh.

I spent half* of yesterday ranting because my western blots didn't work and what I'm doing is essentially exactly the same as the blots I did during my honors year (undergrad - 3 years ago) except with different isolates of the bacteria. So why the hell can't I get it to work now?

I just worked out (from going through my old lab books) that the reason it isn't working now is that I've actually improved my technique. I try to load a particular concentration of cells to the SDS-PAGE gels so that the amount of total protein in each lane is the same. I've been trying to load the same amount of cells now that I was then, only back then I wasn't so good at getting that right so I had up to 4 or 5 times as much protein on the gels than I do now and when I did get the amount right the blots didn't work - too little of the specific protein - only I thought at the time I was stuffing it up the other way. So it's easy to fix - just load 4 times as much and I should be fine - but it's rather frustrating that the blots didn't work just because I loaded the amount of protein I intended to!

I also got the letter from my uni pointing out that I have to hand in by the end of the year. I knew that already, but it's still scary.



*The other half was spent marking first year chemistry exams. I swear they get dumber every year. This year I marked the question about boiling points. It appears that more than three quarters of the students seem to think that when something boils it "breaks down" (as in, they think that the molecules break up into individual atoms). Some of them think it involves hydrogen bonding with water too. meh.
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    frustrated frustrated