problems with DIG

Sasha Kraev bckraev at
Thu Dec 1 13:34:23 EST 1994

Katherine, I do use my blots several times ( up to 5), but this works with
the following combination ( rather far from recommended by the company)
-Boehringer membrane
-Alkaline transfer+UV crosslink of DNA
-Church/Gilbert hybridization buffer at 60 C
-Engler-Blum detection (Anal.Biochem. 210,235-244 (1993))
-no reuse of reagents ( except for the probe )
-everything made in a roller oven ( no bags)
-stripping in 0.2M NaOH-1% SDS for 15 min. at r.t.
Hope this helps.

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