Is FORMAMIDE necessary for hybridizations?

M. Hertel hippocampus at NOSPAM.gmx.ch
Mon Jan 22 12:04:19 EST 2001


Patrick Lynch wrote:
> 
> Always wanted to ask this - is it best to carry out hybridizations in the
> presence or absence of 50% formamide?
> 
> P.


Hello Patrick,
Formamid reduces the hybridization temperature, so you can do Your RNA
hybridizations at 42 C.
An other advantage for hybridizing RNA in Formamid is that possible
contamination of RNase would be inactive.
Always deionize Your Formamid for RNA use with an ion exchange resin if
it is not absolutely free from ionic contaminants.
Of course the church buffer works also very well for northerns if you
want to avoid formamid. For RPA we use up to 80% Formamid, here you need
only some ul...
Moritz






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