Hey, all,
I've been trying to purify a 1.1 kb fragment from an agarose gel, but I had a peak absorbance at 230 in all my samples (using the QiaQuick gel extraction kit), so I'm pretty sure I have a high amount of guanidine thiocynate contamination.
Everything I've heard on this has had people recommend Na-acetate/alcohol precipitation. Possibly stupid question, but would a PCR purification kit (like the Invitrogen PureLink) work as well?
Thanks in advance!
0
1 reply to this topic
#2
perneseblue
-
- Global Moderators
-
- 530 posts
Unlimited ligation works!
8
Neutral
Neutral
Posted 28 May 2011 - 04:44 PM
You can use two washes of PE buffer. That would remove any Guanidium thiocyanate from the membrane.
A PCR purification kit would work... but it will cost more and you would still need to be careful to remove all the guanidium chloride from the membrane with wash buffer. A guanidium chloride solution is used to make the DNA bind to the column silicon matrix.
NaAcetate and ethanol percipitation, followed by 70% ethanol wash would solve your problems.
A PCR purification kit would work... but it will cost more and you would still need to be careful to remove all the guanidium chloride from the membrane with wash buffer. A guanidium chloride solution is used to make the DNA bind to the column silicon matrix.
NaAcetate and ethanol percipitation, followed by 70% ethanol wash would solve your problems.
May your PCR products be long, your protocols short and your boss on holiday
