RT-PCR on cells

Nicolai Stefan nicolai at uia.ua.ac.be
Thu Sep 12 05:24:04 EST 1996


Hello,

We'd like to perform a semiquantitative PCR. If possible, we would like
to start directly from mammalian cells (rat C6 celline or blood
lymphocytes), to eliminate the need to prepare RNA first. So we'd like
to prepare first strand cDNA on the cells, and use an aliquot of this
reaction for the subsequent PCR reaction.
In the future we also like to use Tth enzyme, to be able to do a single
tube quantitative RT-PCR directly on cell material.
Has anyone experience in RT-PCR on cells?

A next problem would be to start from small tissue samples instead of
cells...

Any hint on this subject would be grately appreciated...

Greetings,
Stefan Nicolai 			email: nicolai at uia.ua.ac.be
Dept. Biochemistry
University of Antwerp
belgium



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