Inquiry about DNA recovery by silica binding
netson at uxmail.ust.hk
Sun Oct 19 12:55:49 EST 1997
I have a few questions about the procedures of silica binding..
I have prepare the silica (for binding DNA)
and resuspended in guanidium thiocyanate. After adding the silica to
solution which contain dissolved agarose, DNA and guanidium thiocyanate
salt, I tap the solution to mix them. Then should I incubate the mixture
in ice or at room temp? Which temperature can give better DNA recovery?
Should I use 50% ethanol with 100mM NaCl, Tris-HCl and EDTA or only 70%
ethanol as the washing solution? Finally, which incubation temperature
45, 55 or 65oC can elute the DNA more effective?
One more question is about the safety of guanidium thiocyanate. I
would like to use silica binding to recovery the DNA in mini-prep. I know
that guanidium thiocyanate can release very toxic gas in acidic solution.
Is it possible that toxic gas released in the procedure (e.g solution
III) of miniprep?
Thanks a lot
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