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 at igbmc.u-strasbg.fr