Chemiluminescent analysis of gene expression on high-density filter arrays

We have optimized conditions for the chemiluminescent analysis of gene expr ession using high-density filter arrays (HDFAs). High sensitivity and speci ficity were achieved by optimizing cDNA probe synthesis, hybridization, and detection parameters. The chemiluminescent expression profile reflected ex pected differences in the transcripts isolated from different sources (plac enta and keratinocytes). We estimated the detection limit for low-abundance message to be 1-15 transcripts per cell, a sensitivity rivaling that repor ted for microarray formats and exceeding that reported for autoradiographic HDFAs. The method allows for short exposure times and reuse of probe. It s hould be equally applicable to techniques such as differential screening of cDNA libraries and differential display PCR.