Help: starting RNase protection assays

Merideth Kelley mlkelley at u.washington.edu
Tue May 23 17:34:47 EST 1995


I need to use RNAse protection to evaluate message levels in transgenic 
mice.  I'm starting from scratch with the chapter from "Current Protocols 
in Mol. Biol." and the Ambion catalog.  Answers to any of the following 
questions will be greatly appreciated.

Is Ambion's RPA-II kit worth the money?  
Does anyone out there have a reliable protocol and/or friendly advice for a 
	beginner?  
Is it worth buying template for size markers?  $75 for 5 ug seems a bit 
	steep.
Does anyone have a plasmid containing a mouse housekeeping gene cDNA that 
	I could use as a probe template for positive control/quantitation?
Who makes the best kit for making probes?  Can I get by with ordering new 
polymerases and using the buffers, rNTPs, etc. that I've used in the past 
for riboprobes? 
Would it be worthwhile to begin by using RNA transcribed in vitro from my 
	transgene constructs?

Thank you!

Merideth Kelley

mlkelley at u.washington.edu



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