IDENTIFICATION AND VERIFICATION OF DIFFERENTIAL DISPLAY CDNAS USING GENE-SPECIFIC PRIMERS AND HYBRIDIZATION ARRAYS

An accurate and streamlined approach to differential display (DD) band identification and verification is described. To minimize false posit ives, the strategy avoids the use of impure Northern blot probes obtai ned from PCR-amplified DD bands. To increase throughput, the cloning o f DD bands is replaced by a gene-specific primer approach, and hybridi zation arrays are rued in place of Northern blots. In summary, DD band s obtained with long primers were directly sequenced to allow the desi gn and synthesis of gene-specific primers, which were then used to PCR -amplify homogeneous probes for the verification of expression pattern s by hybridization array analysis. Differential expression of 60 of th e 63 genes tested was confirmed Thug false positives are not inherent to DD. The results demonstrate the power of DD used with hybridization arrays to rapidly generate information on expression patterns of diff erentially expressed genes.