title: GSM2450184: 160613; Homo sapiens; miRNA-Seq
library_construction_protocol: 200 ng of total RNA from each sample was used for library preparation. The TruSeq Small RNA Sample Prep Kit (Illumina) was used for the first batch, while the NEBNext Multiplex Small RNA Library Prep Set (Illumina) was used for the second and third batches. RNA adapters were ligated to 3’ and 5’ ends of the RNA and the adapter-ligated RNA was reverse transcribed into single-stranded cDNA. The RNA 3’ adapter was designed to target microRNAs and other small RNAs that have a 3’ hydroxyl group resulting from enzymatic cleavage by Dicer or other RNA processing enzymes. The adapter used for the first batch has the following sequence: TGGAATTCTCGGGTGCCAAGG, while the one used for the second and the third batches has the following sequence: AGATCGGAAGAGCACACGTCT. The cDNA was then amplified by PCR, 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. A 0.5% PhiX spike-in was also added in all lanes for quality control. Each multiplexed library was hybridized to one lane of the two 8-lane High-Output single-read flow cells on a cBot Cluster Generation System (Illumina) using TruSeq Single-Read Cluster Kit (Illumina). The clustered flowcell was loaded onto a HiSeq 2000 sequencer for a multiplexed sequencing run which consists of a standard 36-cycle sequencing read with the addition of a 7-cycle index read.
attributes.tag: GEO Accession
attributes.value: GSM2450184