Northern blot re-probing

Sunil Shaw sshaw at nospam.bustoff.bwh.harvard.edu
Wed Feb 25 16:16:05 EST 1998


First, always probe with the standard last. Many standard RNAs (actin, GAP) are so
abundant, that even if stripping is 99% effective, the remaining signal is too
much.

Regarding the failure of actin to reprobe, maybe all the actin RNA hybridized with
probe, and then when the probe dried, the RNA:DNA hybrid got cross linked to the
filter. Now there is no available RNA left. Just a guess, though.







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