dsRNA synthesis

William R. Morgan wmorgan at ACS.WOOSTER.EDU
Thu Jul 9 11:06:15 EST 1998


Dear Worm-Breeders,

Has anyone synthesized both RNA strands in a single reaction?

I'm preparing to synthesize double-stranded (ds) RNA for RNA-mediated
inhibition experiments.  The "RNAi info from the Fire Lab" manual describes
the synthesis of each RNA strand separately, with a subsequent annealing
reaction to produce dsRNA.  I'd like to synthesize both RNA strands
simultaneously in a single reaction using both T3 and T7 RNA polymerases.
Both polymerases work under identical conditions (according to Promega) and
I've cleaved the DNA templates at sites flanking the T7 and T3 promoters
(either, BssHII or PvuII, depending on the pBluescipt vector), so it seems
like it should work.

Has anyone tried simultaneous double-strand RNA synthesis in vitro (or have
any other comments on this)?

If there's sufficient response and interest, I'll post a summary to the group.

Bill Morgan


William R. Morgan
931 College St.
Department of Biology			Phone:  330-263-2026
The College of Wooster                  FAX:    330-263-2378
Wooster, OH  44691                      E-mail: wmorgan at acs.wooster.edu





More information about the Celegans mailing list