hybridization incubators

cjs at canctr.mc.duke.edu cjs at canctr.mc.duke.edu
Tue Feb 18 12:06:22 EST 1992

In article <01GGIUSO3MCG95MQH0 at epavax.rtpnc.epa.gov>, MRM at EPAVAX.RTPNC.EPA.GOV writes:
>We've been attempting to use a Robbins hybridization incubator for
>some time now (glass bottles that rotate).  There seems to an
>enormous amount of variability in the quality of the blots we
>obtain and it seems to be probe and hybridiation fluid dependent.
>Some of the blots have so much background that they cannot be used,
>however, the same probe and fluid hybridized in a "seal-a-meal"
>bag would look great.  We'ev tried different blotting media as
>well with not much success.  Does anyone have a universally
>good protocol for this technique that works with all probes?
>Marc J. Mass, Ph.D.
>MRM at epavax.rtpnc.epa.gov

	We've been using the Robbins incubators for about a year now, and
haven't really encountered the problems which you describe. For library
screens we typically use the NEN Colony/Plaque Screen Filters and
use the Formamide hybridization solution at 42 dC.  For screening 
southerns, we typically use Genescreen plus and the hybridization
solution varies, dependent on the user. I tend to stick with the
Formamide hybridization solution and 42 dC, however several other
members of this lab, use a non-formamide solution and incubate
at 65 dC. The only  problems that we occasionally encounter are
where bubbles arise between the filter and the tube. I will check around
for further information about problems and procedures that other 
members of this lab use. We have 3 incubators here, so I'm not sure
how everyone does there incubations.
		Caryl Schwartzbach

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