GSM3351623: CD34+ BM donor 5; Homo sapiens; RNA-Seq
RNA was processed from young adult healthy CD34+ bone marrow. The total RNA was extracted by using mirVana miRNA Isolation Kit (Lifetech, Grand Island, NY) with total RNA extraction protocol. In brief, freshly prepared cells were immediately lyzed by Lysis/Binding Buffer, treated with Homogenate Additive, and followed by Acid-Phenol:Chloroform extraction according to the standard protocol. The supernatant was mixed with ethanol and passed through Filter Cartridge. The bound RNA was then washed and eluted. The RNA concentration was measured by Nanodrop (Thermo Scientific, Wilmington, DE) and its integrity was determined by Bioanalyzer (Agilent, Santa Clara, CA). Using PrepX mRNA Library kit (WaferGen) and Apollo 324 NGS automatic library prep system, the depleted or isolated RNA was RNase III fragmented, adaptor-ligated and Superscript III reverse transcriptase (Lifetech, Grand Island, NY) converted into cDNA, followed by automatic purification using Agencourt  AMPure XP beads (Beckman Coulter, Indianapolis IN). Next, using the universal (SR) and index-specific primer with limited PCR cycle number (~13), sample-specific index was added to each ligated cDNA sample and the amplified library was enriched by AMPure XP beads purification with the final elution volume of 16 µl.
GEO Accession
GSM3351623