Transcriptome of finger millet- control sample
Library preparation was performed at Genotypic Technology’s Genomics facility following Illumina TruSeq RNA library protocol. 1ug of Total RNA was subjected to Poly A purification of mRNA. Purified mRNA was fragmented for 4 minutes at elevated temperature (94oC) in the presence of divalent cations and reverse transcribed with Superscript III Reverse transcriptase in the presence of First strand mix. Second strand cDNA was synthesized using Second strand mix. 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).