GSM7832789: JI64, Embryo, DevStage 3, replication 2 [E_JI64.3.2]; Pisum sativum subsp. elatius; RNA-Seq
Total RNA was isolated using PureLink™ Plant RNA Reagent (Invitrogen). Residual DNA was removed by Baseline-ZERO DNase (Epicenter) treatment followed by phenol/chloroform extraction. Library preparation started with QC of RNA by Nanodrop, Agarose gel electrophoresis RNA degradation and contamination test and Agilent 2100 integrity checks and quantification. mRNA from organisms was enriched using oligo(dT) beads. Fragmentation of mRNA was done  by fragmentation buffer. The cDNA is synthesized by using mRNA template and random hexamers primer and a custom second-strand synthesis buffer with dNTPs, RNase H and DNA polymerase I. Finally, series of terminal repair, A ligation and sequencing adaptor ligation were performed. cDNA library was completed through size selection and PCR enrichment.