Help stripping non-radioactive northerns

Celedonio Gonzalez cglez at ull.es
Fri Jul 14 08:48:03 EST 1995


Hi everybody,

     I'm doing non-radioactive northerns with the Boheringer
Manheim (BM) system: RNA on Nylon(+) membrane hybridized with a
digoxigenin-labelled probe + anti-digoxigenin antibody-Alkaline
phosphatase + chemoluminiscent substrate for Alkaline Phosphatase
(CDP-star).

     They work OK, but now I want to reprobe the membranes. I've
tried two methods for stripping with NO SUCCESS so far:

     Method 1 (recomended by BM).- Rinse in water, wash 2x 30 min
with 50% dimethylformamide, 1% SDS at 68§C, rinse in water, rinse
in 2xSSC.

      Method 2 (Anal. Biochem. 210: 235).- 5 min in water, 20 min
in 5xSSC, 2 min in 0.1% SDS at 95§C.

     Anybody knows of a protocol that works for this purpose???
Please e-mail me (cglez at ull.es), I'm not suscribed to this
newsgroup. I will post a summary with the answers (if there's any).
Thanks in advance.

Celedonio Gonzalez
Departmento de Bioquimica
Universidad de La Laguna
Tenerife. Canary Islands. SPAIN



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