RNase- or S1 nuclease Protection Assay & ABI377

Arthur Arthur
Wed Aug 27 08:40:05 EST 1997


Does anyone have experience with or is currently trying to set up RNase- 
or S1 nuclease Protection Assays using the ABI377 sequencer? I am 
considering this application as the probe is stable in contrast to the 
radioactive versions of the protection assays. However, I don't have any 
experience with protection assays, thus I may be ignorant of very 
obvious pitfalls. What could be possible problems when switching to 
fluorescent probes? Does RNA polymerase incorporate fluorescent 
nucleotides? Is the fluorescent probe/RNA complex resistent to RNase or 
S1 nuclease?
Thanks in advance for thinking with me.
With best regards,

A.Osterop Ph.D.
Erasmus University Rotterdam
The Netherlands
osterop at tch.fgg.eur.nl



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