454 GS FLX sequencing
Total RNA from Litomosoides sigmodontis adult female was converted to double stranded cDNA using Evrogen's MINT cDNA synthesis kit. First strand cDNA was synthesised using reverse transcriptase (RT) from a 3'-primer with oligo(dT) sequence that annealed to the poly-A stretch of RNA and synthesised cDNA until the 5' end of the mRNA. Finally, double stranded cDNA synthesis was performed using PCR amplification, and the final product contained the same MINT adapter sequence at both 3' and 5' ends. The cDNA was fragmented, size-selected, library-prepped, and sequenced according to standard Roche-454 FLX and Titanium protocols.