cross-hybridization in T7 RNA assay

rla at canctr.mc.duke.edu rla at canctr.mc.duke.edu
Tue Jun 29 08:20:25 EST 1993


We are  picking up a lot of anti-sense msg in vivo and in reconstruction
expts using T7-driven hot RNA. Hybridization to sense DNA is significant (maybe
10% of hybridization to anti-sense DNA).  We've washed these blots at
extraordinarily high stringency (like 0.01x SSC, 0.1% SDS, 68 C) and still have
hybridization to sense DNA (BTW, this is all being done using M13 ssDNA on
filters).

Any comments on how we might reduce this noise?

Rick Alston, Associate Director		rla at canctr.mc.duke.edu
Molecular Biology Database Facility	Voice: 919.684.5232
Duke Comprehensive Cancer Center	FAX: 919.684.5296
DUMC 3424, Durham NC 27710




 



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