Journal
SCIENCE
Volume 316, Issue 5830, Pages 1481-1484Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1137325
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We describe a sensitive mRNA profiling technology, PMAGE ( for polony multiplex analysis of gene expression), which detects messenger RNAs ( mRNAs) as rare as one transcript per three cells. PMAGE incorporates an improved ligation-based method to sequence 14-nucleotide tags derived from individual mRNA molecules. One sequence tag from each mRNA molecule is amplified onto a separate 1-micrometer bead, denoted as a polymerase colony or polony, and about 5 million polonies are arrayed in a flow cell for parallel sequencing. Using PMAGE, we identified early transcriptional changes that preceded pathological manifestations of hypertrophic cardiomyopathy in mice carrying a disease-causing mutation. PMAGE provided a comprehensive profile of cardiac mRNAs, including low-abundance mRNAs encoding signaling molecules and transcription factors that are likely to participate in disease pathogenesis.
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