GSM7233638: Salt_NN24h_L_2; Hordeum vulgare; RNA-Seq
After qualified RNA samples were detected, magnetic beads with Oligo(dT) were used to enrich mRNA of eukaryotes mRNA was divided into fragments by fragmentation buffer. cDNA was synthesized by six-base random hexamers based on mRNA template. Buffer solution, dNTPs, and DNA polymerase I and RNaseH were then added to synthesize the two-strand cDNA, which was then purified using AMPure XP beads. The purified double-stranded cDNA was first end-repaired with A tail and connected with sequencing connectors, and then AMPure XPbeads were used for segment size selection. After that, USER enzyme was used to degrade the second strand of cDNA containing U, so that the final sequencing information came from the first strand cDNA, thus preserving the strand orientation of mRNA. Finally, PCR products were amplified and purified by AMPure XP beads to obtain chain specific cDNA libraries.