Abstract
We have developed an AFLP-based gene expression profiling method called High Coverage Expression Profiling (HiCEP) analysis. Improvements on the selective PCR technique we have reduced the rate of false-positive peaks to approximately 2% and consequently the number of peaks, including overlapping peaks, has been markedly decreased. The low false-positive rate enables us to analyze gene expression with wide coverage by means of 4-nucleotide recognition restriction enzymes for fingerprinting mRNAs. The method detects 70-80% of all transcripts, including non-coding transcripts, unknown and known genes. We can determine the relationship between peaks and original transcripts unequivocally. This will make it practical to prepare a database of all peaks, allowing gene assignment without having to isolate individual peaks. The procedure is highly reproducible and sensitive enough to detect even a 1.2-fold difference in gene expression. The method requires no sequence information and so is applicable even to eukaryotes for which there is no genome information available. [J Radiat Res 44:381 (2003)]
