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Single worm reverse-transcriptase (RT) PCR
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g.Jo...@lshtm.ac.uk
Sep 11, 2009, 7:20:12 AM9/11/09
to cele...@magpie.bio.indiana.edu
Dear colleagues,
I'm advised that putative positives from RNA-interference (RNAi) experiments (using feeding strains from Geneservice {Fraser et al [2000]. Nature 408 (6810) 325-30}) can be checked by RT-PCR. This requires reverse transcriptase reaction on single worms.
Single worm PCR lysis buffer doesn't seem amenable to reverse transcriptase reaction {?}
Does anyone have a method/protocol for single worm RT-PCR ?
G. Joshua.
G.W.P.Joshua
London School of Hygiene and Tropical Medicine
Dept. Infectious and Tropical Diseases,
Keppel St.,
London WC1E 7HT.
Tel: (+44) (0)20 7927 2028
FAX: (+44) (0)20 7637 4314
E-mail: g.jo...@lshtm.ac.uk
I'm advised that putative positives from RNA-interference (RNAi) experiments (using feeding strains from Geneservice {Fraser et al [2000]. Nature 408 (6810) 325-30}) can be checked by RT-PCR. This requires reverse transcriptase reaction on single worms.
Single worm PCR lysis buffer doesn't seem amenable to reverse transcriptase reaction {?}
Does anyone have a method/protocol for single worm RT-PCR ?
G. Joshua.
G.W.P.Joshua
London School of Hygiene and Tropical Medicine
Dept. Infectious and Tropical Diseases,
Keppel St.,
London WC1E 7HT.
Tel: (+44) (0)20 7927 2028
FAX: (+44) (0)20 7637 4314
E-mail: g.jo...@lshtm.ac.uk
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