title: Vitisgraft
design: Massive analysis of 3-cDNA end (MACE) - Poly-adenylated mRNA was isolated from 1 g of the large fraction of total RNA using Dynabeads mRNA Purication Kit. First- and second-strand synthesis of cDNA was performed using SuperScript III First-Strand Synthesis System with modied bar-coded 50-end biotinylated poly-T adapters suitable for the Illumina Hiseq2000 ow cell. Subsequently, the cDNA was fragmented to yield 250 base pair (bp) fragments. The 30-ends of the fragmented cDNA were captured with streptavidin beads, while PCR bias-proof technology TrueQuant was used by ligation of TrueQuant adapters to distinguish PCR copies from original copies. The barcoded samples were sequenced simultaneously in one lane of an Illumina Hiseq2000 with 1100 bps