Summary for "Non-specificity for Primer Extension?"

Dan Diaz bl275 at cleveland.Freenet.Edu
Tue Apr 5 10:59:34 EST 1994


In a previous article, smrodems at students.wisc.edu (Steve Rodems) says:

>I believe actinomycin D is an inhibitor of RNA polymerase II.  This is
>included so that any RNA products synthesized in the tube are derived from
>the RT and not from RNA polymerase.  I don't know where the RNAP II would
>be coming from since most protocols call for guanadinium treatment to
>release RNA which I believe has its way with most proteins.

actinomycin D inhibits second strand synthesis during first strand cDNA
synthesis by reverse transcriptase.  it is typically used with MMLV RT,
whereas pyrophosphate is typically used with AMV RT, as MMLV RT doesnt work
well in the presence of pyrophosphate.

diz



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