GSM1370358: L8_12Plex_A; Homo sapiens; miRNA-Seq
30 mg of whole lung was homogenized with Trizol and then total RNA was extracted using the QiaCube system (Qiagen, Valencia CA). RNA quality was determined by Agilent Bioanalyzer 2100 (Agilent Technologies Inc, Santa Clara, CA) One microgram of total RNA from each sample was used for library preparation with the TruSeq Small RNA Sample Prep Kit (Illumina).  RNA adapters were ligated to 3’ and 5’ end of the RNA molecule and the adapter-ligated RNA was reverse transcribed into single-stranded cDNA. The cDNA was then PCR amplified using a common primer and a primer containing one of 12 index sequences. The introduction of the six-base index tag at the PCR step allowed multiplexed sequencing of different samples in a single lane of a flowcell.  One to twelve of these indexed cDNA libraries were pooled in equal amount and gel purified together.  Each library was hybridized to one lane of the 8-lane single-read flowcell on a cBot Cluster Generation System (Illumina) using TruSeq Single-Read Cluster Kit (Illumina). The clustered flowcell is then loaded onto HiSeq 2000 sequencer for a multiplexed sequencing run that consists of a standard 36-cycle sequencing read with the addition of a 7-cycle index read.  A PhiX library was sequenced in lane 4 and used for calibration.
GEO Accession
GSM1370358