Help with biotin-labeled DNA probes

CrouseJ crousej at aol.com
Sun Nov 27 23:50:11 EST 1994


In article <01HJKYRO3NC200ADLI at Post-Office.UH.EDU>, CPedemonte at UH.EDU
(carlos h. pedemonte) writes:

1.  The presence of biotin does not reduce the efficiency of the probe for
RNA
     any more than it does for DNA....(that is it reduces the Tm only
slightly
     over unbiotinylated probes.
2.  The conditions for probe generation are fine and really should not be
changed
     wrt time and temperature.
3.  It is not necessary to remove the unincorporated biotinylated nts.
4.  Yes use a non charged enhanced nylon such as Magnagraph, Hybond or
      Nylon-1.

You should dot blot your unbiotinylated, denatured DNA that was used to
make the probe and throw this in with your Northerns as a positive
hybridization control.

Have you ever considered switching to a different form of
chemiluminescence such as LumiPhos where the light emission lasts longer
and therefore meaningful long term exposures can be done?

Good Luck and Regards.




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