title: Illumina sequencing of Human liver transcript fragment library
design: This method involves several steps that are designed to convert total RNA into a library of template molecules suitable for high throughput DNA sequencing.  The first step involves purifying the poly-A containing mRNA molecules using poly-T oligo-attached magnetic beads.  Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and a high concentration of random hexamer primers.  This is followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH.  Finally the short cDNA fragments are prepared for sequencing on the Illumina Genome Analyzer using reagents provided in the Genomic DNA Sequencing Sample Prep Kit available with the system.