title: GSM9161731: LY10_2; Bos taurus; RNA-Seq
library_construction_protocol: RNA was extracted from bECs using the SMART-Seq® mRNA HT LP (Takara, #032124) and from bESCs using the RNeasy Mini Kit (Qiagen, #74104), both following the respective manufacturer's protocols. Total RNA was purified to obtain mRNA using poly-T beads. mRNA was fragmented and reverse transcribed into first-strand cDNA with random hexamers and M-MuLV Reverse Transcriptase. Second-strand synthesis was done with DNA Polymerase I and RNase H. DNA ends were blunted and adenylated, followed by ligation of hairpin loop adaptors. cDNA fragments (370–420 bp) were selected using AMPure XP system. PCR amplification was performed with Phusion polymerase, universal primers, and index primers. Final PCR products were purified and library quality assessed on Agilent Bioanalyzer 2100.