454 sequencing of Cochliomyia hominivorax Screwworm fly adult female transcript fragment library
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