title: Transcriptome analysis of antennal glands of Penaeus monodon in response to salinity stress
design: Library preparation was performed at Genotypic Technology’s Genomics facility following  Illumina TruSeq RNA library protocol outlined in “TruSeq RNA Sample Preparation Guide”  (Part # 15008136; Rev. A; Nov 2010). Briefly, 1ug of Total RNA by qubit was enriched for  Poly-A using RNA Purification Beads provided with the kit, enriched RNA was fragmented for  4 minutes at elevated temperature (94oC) in the presence of divalent cations and reverse  transcribed with Superscript III Reverse transcriptase by priming with Random Hexamers.  Second strand cDNA was synthesized in the presence of DNA polymerase I and RnaseH. The  cDNA was cleaned up using Agencourt Ampure XP SPRI beads (Beckman Coulter). Illumina  Adapters were ligated to the cDNA molecules after end repair and addition of “A”- base. SPRI  cleanup was performed after ligation. The library was amplified using 8 cycles of PCR for  enrichment of adapter ligated fragments. The prepared library was quantified using Nanodrop  and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).  (Refer Figure 1 for Bioanalyzer profiles of amplified product ePCR1).