miRNA sequencing of Shanbei White cashmere goat: fetal E120 skin 3
After the RNA sample was qualified, sequencing library was generated using NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB, USA.) following manufacturers recommendations and index codes were added to attribute sequences. 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. DNA fragments corresponding to 140~160 bp were recovered and dissolved in 8 L elution buffer. At last, library quality was assessed on the Agilent Bioanalyzer 2100 system using DNA High Sensitivity Chips. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq SR Cluster Kit v3-cBot-HS (Illumia). After cluster generation, the library preparations were sequenced on an Illumina Hiseq 2500 platform and 50bp single-end reads were generated.
E120_3