callus of DH201-2, replicate 3
Stem and callus tissues (100 mg) were collected for the total RNA isolation using the TRIzol reagent (Invitrogen) according to the manufacturers protocol. Then, the quantity and quality of the total RNA were determined by the Agilent 2100 Bioanalyzer and equal amount of total RNA (1 g) was to construct the libraries for BGISEQ-500 RNA-Seq. Briefly, magnetic oligo (dT) beads were used to enrich the ploy(A) mRNAs, which were then fragmented into ~200 bp. Then, the fragments were used for the dscDNA library construction by random hexamer (N6) primers, followed by the end repair using phosphate at the 5 end and sticky A at the 3 end. Next, sequencing primers were ligated to build the final cDNA library, which was then sequenced on the BGISEQ-500 RS platform at BGI-Shenzhen with paired-end 150 strategy.