RNase protection assays...
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.
Petawawa National Forestry Institute
brutledge at pnfi.forestry.ca
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