Calculating Tm for mismatch on Northern
Mark D. Garfinkel
mg16 at kimbark.uchicago.edu
Thu Jul 27 10:42:28 EST 1995
In article <1995Jul24.133759.8909 at mbcf>, <neale at mbcf.stjude.org> wrote:
>We have a partial cDNA clone from mouse that we wish to screen *human* northern
>blots.
>My question is whether you can calculate a lower stringency and wash of these
>blots the same as you would for Southerns?
Same formula, different effective Tm. It's the difference between
detecting DNA-DNA hybrids and DNA-RNA hybrids, the latter being
more stable, and the differential effects thereon caused by
formamide-containing hybridization solutions. See, for example,
White & Davis (1976) PNAS and Casey & Davidson (1977) NAR for the
original experiments.
Mark
--
Mark D. Garfinkel (e-mail: mg16 at midway.uchicago.edu)
(c) 1995; all rights reserved. Permission granted for Usenet quotation
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