cDNA RDA

SchatzD2 Lab SchatzD2_Lab at QUICKMAIL.CIS.YALE.EDU
Sat Nov 26 17:45:55 EST 1994


                      Subject:                              Time:  5:26 PM
  OFFICE MEMO         cDNA RDA                              Date:  11/26/94

I would like to post the following method on the mol biol methods bulletin
board.
We have adapted the PCR-coupled subtractive hybridization process of genomic
RDA (representational difference analysis)(Lisitsyn  et al. Science 1993 259,
946) for use with cDNA. The technique is fast (approx 3 weeks), reliable, and
very sensitive. We have been able to amplify differentially expressed messages
from both cell culture and tissue sources, with very little background of
false positives (less than 25%). In addition  we were able to compete out
unwanted messages, and by varying the stringency of subtraction, to detect
up-regulated messages. The method has significant advantages over both
traditional subtractive approaches and differential display PCR. Our paper is
currently in Press at NAR, and we would like as many people as possible to try
it. Consequently, if you would like a detailed protocol, please send me (Mike
Hubank) an e-mail message to schatzd2_lab at quickmail.yale.edu telling me who
you are, and on what system you would like to try the method. I shall be happy
to help with any questions or problems.

Mike Hubank
David Schatz

HHMI Immunobiology
Yale University
New Haven CT 06510





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