S Blots: Alkali or SSC

DYDM6025 at iruccvax.ucc.ie DYDM6025 at iruccvax.ucc.ie
Wed Jun 21 09:10:48 EST 1995


Im planning to do some human RFLP analysis. The probes I plan to use
should detect alleles at 10.5,5.5 ; 2.9,1.5
I plan to use the membrane for both probes , using SDS to remove probes.

Questions:  In the Amersham N + with the alkali transfer they depurinate
	wash in dH20 and then transfer.  Is there a need to denture the
gel. Must you neutralise the membrane afterwards.

The hybridisations are going to be carried out in a plastic container,
so any ideas on hyb solutions from people who are using these types of 
containers.
Regards
Sean



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