[FUN] Dumbest lab experience

Edgar Valencia Morales edgar at cifn.unam.mx
Fri Dec 15 12:45:45 EST 2000


Hi
heres my contribution

I had to design PCR oligos with the EcoRI site to allow me to clone it. well,
the sequence at the site where I wanted to design the oligo had 5 out of 6 of
the bases of the EcoRI site, so, I just had to modify 1 base, then add 3 more
bases to allow the enzyme to cut and everything should be perfect....except
for the fact that "the site" was in the 3'->5' orientation. I saw this at the
begining, but then I thought, the site will be on the other chain too, so
shouldn't be any problem right? well, i always got a product of the right
size which of course I couldn't clone...and of course in the meantime I drove
crazy half of the lab cause I never talk about this site being in the
opposite direction, it was soooo obvious that it should work....til I found a
paper about the mechanism of cutting of EcoRI...

And here is my friend's contribution

he has been doing lot of minipreps to isolate plasmid DNA, always perfect
ok?, but he had never extracted Genomic DNA, so when he did it, he saw the
"smear" in the lane and thought he did something wrong, so, he did it again,
and exactly the same result, I think he should've done it 15 times at least,
always with the same result...until his tutor saw the gel and told him that
was exactly what he must see.

Edgar


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