Can't precipitate DNA!

atgcpaul at yahoo.com atgcpaul at yahoo.com
Sat Oct 25 23:17:46 EST 2003


David Micklem <drmicklem at nospam.com> wrote:

: Wot? No ethanol? Or is it added with the NaAc?

   Ice cold 200 proof ethanol added to PCR with sodium acetate--all
separate solutions.


: Could it be that you are squirting in the 70% ethanol more vigourously
: than the others and dislodging the pellet? I never think they stick as
: well after being dislodged, even if you spin again.

   Since this began, I have been extra careful with the 70%.  It's 70% 
made from 95% denatured ethanol--also works for sequencing rxns.  In my 
last attempt, I added the 70% right on top of the other mix without any 
dumping.  Even if I didn't wash away all the salt, the pellets weren't hit 
directly with 70% and I should have still had some product.

   I think next time I'll switch plates or try different sodium acetate 
mix.



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