Use of labeled primers for differential display

Abstract

Two artifacts introduced in using differential display technology are (1) random priming from dT present from afEnily purification of PolyA+ RNA and (2) hybridization of the a r b i t r q primer to template target sequences on both cDNA strands. We have developed a method eliminating both problems. By separately using 5'-end-labeled (T)&Y and arbitrary primers to label bands and comparing two differential display patterns, we can detect only those products incorporating the (T) l T primer on the 3' ends and the a r b i t r q primer on 5' ends. Those bands that are generated randomly in the PCR are readily detectable and can be ignored.