Wednesday, July 21, 2004

world's. biggest. dumbass. 

So yesterday I waited and waited for the radiation guy (who does have a full head of hair, by the way) and he never came.  The lab next door had just gotten in a shipment of the same stuff the day before, and they said I could use their stuff, and that they would use ours next time they needed it.  (this is against the rules, but screw the rules!  living on the edge, sharing radioactivity, baby!) 

So I made my probe, used the new buffer from the JCL lab, got a little over 50% incorporation (kick-ass!) added to the hyb tubes with my blot and left it.  My beautiful blots will be ready tomorrow, yay!

Last night, I dreamt that Martha Stewart was going to start working in our lab, and that I was going to have to work with her.  I was in a panic, because I knew she was going to be difficult to work with.
She had me chopping onions for my experiment and kept criticizing me about it.  Nothing I did was right.
Then she asked me if I'd boiled my probe.

This was around 3am.  I sat bolt upright in bed.  I didnt' boil my probe before adding it!!!!

Shit.  See - a probe is supposed to be composed of single stranded DNA (think of a ladder, split in half down the middle).  It doesn't like this, so it binds to the DNA you have imbedded on a membrane (the blot).  Since it's radioactive, where ever my DNA of interest is on the blot, that area will become radioactive.  But when you make your probe, it is double stranded (a ladder).  You need to boil it for a minute to break the strands apart, then put in on ice so the strands don't have time to find their complementary partners and just sort of sick together half-assed, leaving alot of single stranded DNA for your probe.

i didn't boil/ice my probe.  I am such a dumbass.

I essentially gave my perfect, wonderful, fabulous blots a 65 degree radioactive overnight bath.  Crap.

So in an attempt to save my wonderful blots, I made yet more probe today, BOILED IT, ICED IT, and added it to the mix (accidentally injecting my pipet tip into one of them.  oy!).  Hopefully this will save my experiment.

If not, I'm quitting science and opening up a soup and sandwhich buisness.  Who's with me???