RNase protection assays...

BOB RUTLEDGE BRUTLEDGE at AM.PNFI.FORESTRY.CA
Mon May 10 16:19:00 EST 1993


Daivid (haviland at kids.wustl.edu) asked about eliminating 
proteinase-K and SDS from the RNase protection assay.  

Two recent BioTechniques articles (BioTech. 13, 847 & 13, 852) 
describe using guanidinium thiocyanate and isopropoanol in a 
rapid RPA protocol, replacing the standard Pro-K/SDS 
inactivation of RNases.  Although we still consider ourselves 
novices of RPA, this seems to be a simple and effective 
alternative to the standard protocol

The first article also describes using RNase T2 in place of 
RNase A/T1.  However, as outlined in a recent posting to BioSci 
M&M, we have found that T2 can rapidly become inactive during 
hybrid digestion.  We have therefore returned to using A/T1.  

Any additional information/experence concerning this alternative 
protocol would be appreciated. 

Bob Rutledge
Petawawa National Forestry Institute
brutledge at pnfi.forestry.ca




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