NORTHERN BLOTS

kvernick at helix.nih.gov kvernick at helix.nih.gov
Tue Sep 6 20:53:07 EST 1994


It sounds like no RNA is on your membrane, by the rDNA probe result. Did you try staining 
the membrane after transfer with methylene blue? THis will show you transfer efficiency. 
I find the best transfer buffer is the one from the Current Protocols Redbook: 2x10min 
50mN NaOH/10mM NaCl (to fragment RNA a little to better leave the gel); 2x8min 0.1M 
Tris-Cl pH 8; 2x15min 20xSSPE; transfer with 20xSSPE



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