non-specific bands in NTC

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Sun Sep 19 04:31:26 EST 2010


Dear Magda,

might be that your polymerase contains traces of DNA from the host it
was produced in. To confirm, add some DNAse to an aliquot of your
mastermix, incubate say 1hr at 37 degC (or overnight if you like) and
then kill the DNAse by heating to 95degC for say 10 minutes (longe is
not so critical, but polymerase of course will decrease in activity).
Then redo your assay. Include a control that demonstrates that DNAse
is really gone; a control plasmid in a reasonable concentration should
be amplified.

Alternatively/additinally, you might try to switch polymerase batch/
brand/type, too.

Hope that helps,

Wo


More information about the Methods mailing list