title: GSM2209694: EVD patient L05_miRNA; Homo sapiens; miRNA-Seq
library_construction_protocol: Whole blood was preserved in Trizol reagent (Ambion). RNA isolation was performed using the miRNeasy mini Kit (Qiagen Cat.No 217004) Sequencing libraries were generated using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB, USA.) following manufactures’s recommendations and index codes were added to attribute sequences to each sample. Briefly, NEB 3’ SR Adaptor was directly, and specifically legated to 3’ end of miRNA, siRNA and piRNA. After the 3’ ligation reaction, the SR RT Primer hybridized to the excess oof 3’ SR Adaptor and transformed the single-stranded DNA adaptor into a double-stranded DNA molecule. Then first strand cDNA was synthesized using M-MuLV Reverse Transcriptase (RNase H-). PCR amplification was performed using LongAmp Taq 2X Master Mix, SR Primer for illumina and index (X) primer. PCR products were purified on an 8% polyacrylamide gel (100V, 80min). DNA fragments corresponding to 140-160bp were recovered and dissolved in 8 μL elution buffer. At last, library quality was assessed on the Agilent Bioanalyzer 2100 system using DNA High Sensitivity Chips, the library preparations was sequenced on an Illumina Hiseq 2500 platform and 50bp single-end reads were generated.
attributes.tag: GEO Accession
attributes.value: GSM2209694