RNA protection assays

Marianna Max drmax at casbah.acns.nwu.edu
Fri Apr 1 16:58:13 EST 1994


In article <hmhz-300394141317 at dent-226-97.dentistry.umich.edu>,
Richard H. Smoke <hmhz at umich.edu> wrote:
>  I am about to begin a project involving RNA protection assays and I am
>going to use an Ambion kit for both the assay and the invitro
>transcription. I have been told to gel purify the the invitro transcription
>but could I use a column instead? It seems it would be alot easier. Any
>suggestions will be appreciated. I am provided with the probes but I have
>to make the RNA myself. Feel free to email me.
>
>Steve Coon
>sdcoon at umich.edu
>University of Michigan Medical Center


Are you talking about gel purifying your hot RNA probe? If so, the idea
behind gel purifying a RNAse protection probe it so that you only include
full length, undegraded probe. How would a column guard against inclusion
of partial length probe? The column is fine for seperating unincorporated
hot nucleotides but those aren't a problem anyway.

Also be sure that the DNA that you use as a template has been fully
linearized and phenol/chloroform extracted.

Max



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