HELP! Lane background in Northern's
bbecker at iastate.edu
Sun Nov 27 15:56:41 EST 1994
In article <Nov24.180359.37000 at acs.ucalgary.ca>,
Lauri Lintott <llintott at acs.ucalgary.ca> wrote:
>Gil (kgilbert at cmp.hmc.psu.edu) wrote:
>: Here are some more specifics which may help someone with my background
>: I run a 1.2% agarose/0.4M formaldehyde gel; downward capillary transfer
>: in 10X SSPE overnight; membrane is washed in 5X SSPE @ 60 degrees for 5
>: min, then baked @80 degrees for 1 hr, and UV crosslinked in a
>: Stratalinker (200 ujX100).
>: After O/N hybridization, washes are as follows:
>: 1. Quick rinse in 2X SSPE
>: 2. 2 x 15 min washes in 2XSSPE/0.5%SDS at hybridization temperature
Are you sure your cDNA probe does not contain long stretches of AAAA
which would result in hybridization to all mRNAs with a poly A-tail?
Just a thought ...
Email: bbecker at iastate.edu
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