RNA-Seq of root of Allium mongolicum
Total RNA was isolated using Trizol reagent (Life Technologies, lnc., Gaithersburg, MD) according to the manufacturers protocol. Extracted total RNA was quantified with a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and qualified by agarose gel electrophoresis. cDNA library construction and sequencing were conducted by Vazyme Biotech Co., Ltd (Vazyme Biotech, Nanjing, China). In brief, total RNA was first purified with RNase free DNase, enriched with mRNA Capture Beads (Vazyme Biotech, Nanjing, China) and fragmented. cDNA was synthesized using random hexamer-primers, taking fragmented mRNA as templates. Double-strand cDNA was then purified with VAHTSTM DNA Clean Beads (Vazyme Biotech, Nanjing, China), end repaired, added A-tail and sequencing adaptor. Proper fragments were selected, PCR amplified and purified to create the final cDNA library. Finally, the library was sequenced using the illumina HiSeqTM 2000 at Vazyme Biotech Co., Ltd, China. Before assembly, Raw reads with adaptors and unidentified nucleotides, as well as low quality reads were first filtered. Clean reads were then assembled to obtain unigenes using the Trinity software as described