miRNA Gene 454 Sequencing, AML
Coordinates for 695 known human miRNA genes were obtained from MiRbase release 12.0. Allowing for approximately 100 bp of flanking sequence, 1,378 PCR amplicons were designed using in-house primer design algorithms.  These were later supplemented with 18 amplicons from low-coverage regions, bringing the total to 1,396 amplicons (average size: 315 bp).  PCR reactions were performed separately to produce 1,396 products from whole genome amplified (WGA) tumor genomic DNA of patient 933124.  All PCR products (3 *L each) were pooled together and sequenced on * plate of the 454-XLR platform (Roche).