Deep-transcriptome and ribonome sequencing redefines the molecular networks of pluripotency and the extracellular space in human Embryonic Stem cells. Homo sapiens

Cytoplasmic polyadenylated RNA from human embryonic stem cells (HES2) was separated on a 1.2% agarose and gel slices were excised corresponding to the following sizes: 0 to 0.5 kb; 0.5-2 kb; 2-3.5 kb; 3.5-6.5 kb and 6.5-20+ kb. Slices were dissolved and extracted RNA (1% of each fraction) was run on the Agilent Bioanalyser (Agilent) to confirm the correct size distribution and yield. Library molecules were clonally amplified onto one micron magnetic beads according to the SOLiD Template Bead Preparation protocol and sequenced using a SOLiD Analyzer according to manufacturer's instructions. Overall design: Human ES cells size fractionated mRNA