Help: stripping northern blots

h-seifert at nwu.edu h-seifert at nwu.edu
Mon Jan 31 15:36:56 EST 1994


In article <2i3do5$97r at gazette.bcm.tmc.edu> Jorges at bcm.tmc.edu (Jorge L. Sepulveda) writes:
>Hi, netters,
>
>I have been using Zeta-probe nylon membranes and pre-hyb/hybridizing at
>65¡C in a solution containing 7% SDS. I get very high signal to noise
>ratio with no background at all. The problem is that I can't stripp the
>probes (boiling in water for 20' or heating overnight at 65¡C in 80%
>formamide hardly knocks off any counts from the membrane). And yes, I
>don't let the membrane dry at any time. The probes are not even homologous
>(e.g. human GAPDH for rat RNA).
>
>Thanks for any clues.

Wash the blot twice in water to remove salts and SDS.

Hank Seifert

>
>
>Jorge L. Sepulveda,
>
>Dept. of Pathology,                        Email: Jorges at bcm.tmc.edu
>Baylor College of Medicine                 Telephone: 713-798-7330
>One Baylor Plaza, Houston, TX 77030        Telefax: 713-798-5838



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