NEW PRIMER STRATEGY IMPROVES PRECISION OF DIFFERENTIAL DISPLAY

To increase the reproducibility and to reduce the false positives in t he initial mRNA differential display, modified long composite primers were developed based on both mRNA differential display and RNA arbitra rily primed PCR fingerprinting methods. Ten-base nucleotides were adde d at the 5' ends of the primers used in the initial mRNA differential display. These included a restriction site to aid cloning. PCR began w ith one low-stringency cycle (40 degrees C for annealing) followed by 35 high-stringency cycles (60 degrees C for annealing). The mollified method significantly improved the reproducibility and sensitivity of t he mRNA differential display while still keeping the characteristics o f the original method.