Guanidine-CsCl RNA microprep

Fri Jul 10 16:00:07 EST 1992

  For many years we have used successfully the method of Chirgwin and coworkers
to prepare RNA from cultures of 10,000,000 to 100,000,000 lymphocytes
(Forsdyke, 1984. Can.J.Biochem. 62, 859-864). RNA was separated from other
macromolecules in a 5ml tube in a Beckman SW 50.1 rotor. Recently we tried to
scale this down for 1,000,000 lymphocytes. We followed the procedure described
by Brenner and workers (Biotechniques 7, 1096-1103). This employs the Beckman
TL-100 bench-top ultracentrifuge and samples (0.2ml) are spun in an angle-head
TLA-100 rotor for 2h at 80,000 r.p.m. Our recoveries of RNA were poor. We
wonder if anyone out there has any information or references which might guide
our troubleshooting of the method?
                                  Sincerely,   Don Forsdyke, Queen's University
                                                             Kingston, Canada.

More information about the Methods mailing list