Briefly, 1 of total RNA was reverse transcribed into cDNA
RT and direct DNA sequencingThe presence of the TMPRSS2:ERG fusion gene was established using RT for all 165 prostate cancer patient samples, described elsewhere (Nam et al, 2007). Briefly, 1 of total RNA was reverse transcribed into cDNA using QuantiTect Reverse Transcription Kit (Qiagen Inc., Valencia, CA, USA) in the presence of random and oligo dT primers. All reactions were performed with both the forward and reverse primer sets (F TAGGCGCGAGCTAAGCAGGAG, R GTAGGCACACTCAAACAACGACTGG) that yields a 125 bp and (F CAGGAGGCGGAGGCGGA, R GGCGGTTGTAGCTGGGGGTGAG) that yields a 595 bp product as described by Tomlins et al (2005).
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