detergents for single cell RT-PCR

Simon Whitfield simon at igbmc.u-strasbg.fr
Fri Mar 19 13:59:01 EST 1999


I am trying to optimise conditions for single cell RT-PCR. The tests I
have done seem to suggest that the RT step is critical.
Currently I am lysing the cells in RT mix containing 0.1% NP40 but
I am not convinced that this is reliably freeing up the RNA for reverse
transcription. Does anyone know any detergents (e.g. Triton X100, Tween
etc) and/or concentrations of detergents that will not inhibit the
MMLV-RTase or the Taq but will do a good job of lysing the cells and
dissociating all of the ribonucleoprotein complexes?

Thanks for your help,
Simon

simon at igbmc.u-strasbg.fr



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