Illumina HiSeq 2000 paired end sequencing
Trancriptome sequencing of O. algarvensis worms for gene detection
From the total RNA sample poly(A)+ RNA was isolated which was fragmented with ultrasound (2 pulse of 30 sec at 4°C). First-strand cDNA synthesis was primed with a N6 randomized primer. Then Illumina TruSeq adapters were ligated to the 5' and 3' ends of the cDNA. The cDNA was finally amplified with PCR using a proof reading enzyme. For Illumina sequencing, the cDNA in the size range of 300 –500 bp was eluted from a preparative agarose gel.