RNA Seq of alfalfa: 250 mM NaCl for 6 h, replicate 1
In brief, total RNA was purified to enrich poly (A) messenger RNA (mRNA) using magnetic oligo (dT) beads. The target RNA was fragmented and used for double-stranded cDNA (dscDNA) library construction by random hexamer (N6) primers. End repaired the dscDNA to phosphate at 5 end and stickiness A at 3 end, then ligated the adapter with stickiness T at the 3 end of the dscDNA. Two specific primers were used to amplify the ligation product. Finally, denature the PCR product by heat and the single strand DNA was cyclized by splint oligo and DNA ligase. The sequence library was constructed and sequenced on the BGISEQ-500 RS platform at BGI Shenzhen.