3' RACE-PCRs for small rRNAs

N. Salazar nsalazar at U.WASHINGTON.EDU
Sat Apr 26 16:00:36 EST 1997


Hi there,
		I am trying to get the 3' end sequences for the P.
falciparum small rRNAs but I am not getting the expected PCR amplified
bands (between 50 and 120 bp in size).

I am using total RNA which is C-tailed. Then I produce cDNA with an
oligodG, and this cDNA is used for the PCR reaction, together with a gene
specific primer for each individual rRNA. Any suggestions are really
appreciated it, to see whether I can get the expected bands. I have
already tried changing the annealing temperature (between 46 up to 50C)
and also changing the Mg++ concentration (between 0.5 and 3.0 mM) but I
get the same negative results.

		Thanx,
			Nelson Salazar
			nsalazar at u.washington.edu




More information about the Methods mailing list