Efficient targeted transcript discovery via array-based normalization of RACE libraries
作者： Catherine Ucla , Carine Wyss , Jorg Drenkow , Erica Dumais , Ryan R Murray
DOI: 10.1038/NMETH.1216
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摘要: Rapid amplification of cDNA ends (RACE) is a widely used approach for transcript identification. Random clone selection from the RACE mixture, however, an ineffective sampling strategy if dynamic range abundances large. To improve efficiency human transcripts, we hybridized products reaction onto tiling arrays and detected exons to delineate series reverse-transcriptase (RT)-PCRs, through which original population was segregated into simpler populations. We independently cloned sequenced randomly selected clones. This approach, RACEarray, superior direct cloning sequencing because it specifically targets new transcripts often results in overall normalization abundance. show theoretically experimentally that this leads indeed efficient investigated multiplexing by pooling reactions multiple interrogated loci before hybridization.
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