title: Artemisia annua
design: Library preparation was performed at Genotypic Technology’s Genomics facility following IlluminaTruSeq RNA library protocol outlined in “TruSeq RNA Sample Preparation Guide” (Part # 15008136;Rev. A; Nov 2010). 1ug of Total RNA was subjected to Poly A purification of mRNA. Purified mRNAwas fragmented for 2 minutes at elevated temperature (94 o C) in the presence of divalent cations andreverse 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 cDNAwas cleaned up using Agencourt Ampure XP SPRI beads (Beckman Coulter). Illumina Adapters wereligated to the cDNA molecules after end repair and addition of A base. SPRI cleanup was performedafter ligation. The library was amplified using 8 cycles of PCR for enrichment of adapter ligatedfragments. The prepared library was quantified using Nanodrop and validated for quality by running analiquot on High Sensitivity Bioanalyzer Chip (Agilent). (Refer Figure 1 for Bioanalyzer profiles ofamplified product ePCR1)