Transcriptome-seq of Sinowilsonia henryi
Total RNAs were isolated from young leaves using a CTAB procedure (Chang et al., 1993). mRNA was purified with the RNA Clean-up Kit (Tiangen, Beijing, China) according to the manufacturerĄŻs instructions. The purified RNA was subsequently fragmented into small pieces (200 bp) by the fragmentation buffer. Then, the cleaved RNA fragments were used for first-strand cDNA synthesis using reverse transcriptase (Invitrogen, USA) with random hexamer primers. Subsequently, second-strand cDNA was synthesized using RNase H and DNA polymerase I (New England BioLabs). Illumina paired-end sequencing adapters were then ligated to the ends of the 3ĄŻ-adenylated cDNA fragments. The cDNA library was sequenced by on Shanghai Haiyu Biotechnology Co., Ltd. on the Illumina HiSeq 2000 instrument. Before assembly, raw reads were filtered to remove adaptor sequences, low-quality reads and reads containing poly-N. Transcriptome assembly was performed using Trinity package (version2013-02-25).