3'-END CDNA POOL SUITABLE FOR DIFFERENTIAL DISPLAY FROM A SMALL NUMBER OF CELLS

We have generated a 3' cDNA pool from the RNA of only 1000 or fewer ce lls by reverse transcription (RT) from an extended oligo(dT) primer wi th a 3' degenerate base and a second strand primer with four degenerat e 3' bases, followed by PCR. Reproducible differential displays (DD) c an be made from this essentially inexhaustible source of DNA. The meth od produced DD patterns that are comparable but not identical in band number and size distribution with those obtained by the original RT-DD technique. Northern blots performed with the excised bands verified a ltered gene expressions. The data indicate that this St-end cDNA pool can supplement current PCR-based methods of expression genetics. This pool of cDNA sequences also provides a reliable source for primer-spec ific gene amplifications.