DNA control on Northern?

Frank Maiwald maiwald at mpiz-koeln.mpg.de
Mon Nov 16 04:29:08 EST 1998


Dear all,

I would like to use a fragment of lambda DNA (part of a cloned insert)
as a positive control in a Northern blot. In my first attempt this
*positive* control did not give any signal, although the target DNA
quantity should be more than enough. I denatured the DNA at 95 C for 10
minutes ice shocked it and loaded it with RNA loading buffer on a
formaldehyde/ MOPS/ agarose gel.
Is it possible that the DNA renatures in the loading buffer or gel? Is
is degraded? Or does my result simply mean what you think at first sight
- the probe does not hybridize to the control?

Any thoughts or hints are welcome; thanks in advance,

Frank



More information about the Methods mailing list