Protocol Online logo
Top : Forum Archives: : Molecular Biology

DNA annealing help - (Jul/09/2008 )

To anneal radiolabelled DNA structures, I put the oligos at 96 degrees then switch off the power to the heat block so they cool slowly overnight.

I have important samples and I have forgot to switch off the heat block so my oligos have been at 96 degrees O/N

What I want to know is - can they be rescued, obviously they can be over denatured but im worried they may have been degraded.

Thanks in advance

-thewhorule2002-

QUOTE (thewhorule2002 @ Jul 9 2008, 09:25 AM)
To anneal radiolabelled DNA structures, I put the oligos at 96 degrees then switch off the power to the heat block so they cool slowly overnight.

I have important samples and I have forgot to switch off the heat block so my oligos have been at 96 degrees O/N

What I want to know is - can they be rescued, obviously they can be over denatured but im worried they may have been degraded.

Thanks in advance


Just bumping this for some help,

cheers

-thewhorule2002-

96 degrees O/N? I'd say they are well and truly denatured! Seriously though, they should be OK. What buffer have you been using for the annealing?
Only 1 way to see: try to use the primer in an expt. Run a gel and see what you get. If it looks OK, go for it.

Is it such a problem to repeat the annealing?

-swanny-