how to set up the ligation - (Jan/25/2011 )

I found out the "trick" I was looking for. It wasn't on the forum but on a protocol I have .
I thought I'd share. It's about a ligation between genomic DNA and adaptors in the presence of PEG. Since PEG is highly viscous, the authors recommend to vortex the master mix before adding the ligase and then mix gently after adding it. Didn't know ligase was fragile.

Hi Pito
Sorry for delay..been busy .
Yes, i clean up as much as I can. usually my A260/A280 is close to 1.8. As for the adaptors..no they don't say. I think I'll figure it out. I was annoyed this week because my amplified library has tons of primer dimers. Then yesterday, I found the recovery gels from Lonza and that made my day .
We are using flashgels already so I can't wait to try these guys out. Is anyone using it?

(We are not university and not biotech...we are contractors for the DoD).

Ah the PEG trick, yes, you are right there.

DoD?
Is that the department of defense?
Are you doing research on killed soldiers? uh?

How did you guess?

Maddie on Mon Feb 7 23:19:28 2011 said:

I was in the army once...

not american army anyway but for some reason it did ring a bell. No idea why really.
So you are working on soldiers then? or do they research all kind of "corpses"?