RNA-Seq of Prunus salicina A1
Using these short fragments as templates, random hexamer-primers were used to synthesise first-strand cDNA. Second-strand cDNA was synthesised using buffer, dNTPs, RNaseH and DNA polymerase I. Sequencing adapters were ligated to the short fragments after purification using a QIAquick Gel Extraction Kit. The adapters were used to distinguish different sequencing samples. Fragments of 200 ± 25 bp were then separated by agarose gel electrophoresis and used as sequencing templates for polymerase chain reaction (PCR) amplification. Finally, the cDNA library was sequenced using an Illumina HiSeq 2000 system.