title: 454 sequencing of Cochliomyia hominivorax Screwworm Fly larvae transcript fragment library
design: PolyA RNA was isolated from total RNA using MicroPolyA Purist Kit (Ambion Inc.).  PolyA RNA was used to synthesize blunt-ended double stranded cDNA using the Just cDNA Double-Stranded cDNA Synthesis Kit (Stratagene Inc., La Jolla, CA) and the supplied random primers.  DNA sample preparation for sequencing on the 454/Roche GS_FLX was performed as described by the manufacturer.  Briefly, this entailed shearing the DNA via nebulization and subsequent end repair, followed by ligation of adapter sequences and a second round of end repair to yield a blunt ended DNA ibrary that is quantified and diluted prior to amplification via emPCR.  After emPCR enrichment, the DNAs were loaded onto a 454 Roche GE-FLX for massively parallel pyrosequencing