If you forget to change the flourophore [spell check wants it to be froufrou now wouldn't that be more fun?] to the one you will use in the RT-PCR protocol that the machine uses as a default setting, it doesn't matter what you did because the whole thing is read wrong and no, it can not be changed afterwards.
Duh.
That's when you redo things.... and then redo things... (because you forgot something else, or missed something small but since it is real time quantitative PCR it is sensitive... and yeah... sloppy you...) Just button up and redo it. Try not to shake when you pipette the small (minuscule) volumes and make 1000+ dilution series (OK, I might be over exaggerating just a bit).
I ended up designing new primers for sequencing... since it came back "wacko-we-don't-want-to-bind" last time again. Now, I have a new batch of purified fragments (just the idea that I am sequencing PCR fragments makes my tummy cringe but that's what they want....and truly, it makes it less impossible but still...) and should probably send them in with the "semi-old" primers anyway since it doesn't hurt to try - and hope is the last thing that dies, right?!
On the flip side I guess, I've been sort of the boss all this week since my boss is gone on vacation. It's interesting being the newest hire but being sort of in charge. Even if it doesn't increase my pay, or is visible in my job description... really, mostly it's about taking responsibility for things (and do more stuff for others), but it has felt a bit fun.
Now, time to leave soon. Just need to finish that sequence order...
1 comment:
Oh that sucks.
B.T.W. my favorite thing spell check does is suggest that electroporate be changed to electro orate.
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