Lycoris aurea Transcriptome Sequencing
Total RNA was extracted from these materials using TRIzol Reagent (Invitrogen, USA) according to the manufacturer’s instructions. The quality of total RNA was determined using a NanoDrop spectrophotometer (Thermo, USA) and RNA samples with a 260 of 280 ratio from 1.9 to 2.1were selected for the next analysis. After that, equal quantities of total RNA from each sample (~0.35 mg total RNA) were mixed together. The cDNA library was constructed using the CreatorTM SMARTTM cDNA library construction kit (Clontech Laboratories Inc., Mountain View, CA, USA) and following the manufacturer’s protocol step-by-step. With agarose gel electrophoresis and extraction of DNA from gels, DNA bands (500~800 bp) were purified, blunt ended followed by ligation with adapters and finally immobilized on beads. The quality control of a double DNA library was performed using High Sensitivity Chip (Agilent Technologies). The concentration and the proper ligation of the adapters were examined by using TBS 380 Fluorometer. After the examination, one-plate, whole-run sequencing was performed on Roche 454 GS FLX Titanium chemistry (Roche Diagnostics, Indianapolis, IN, USA) by Shanghai Majorbio Bio-pharm Biotechnology Co., Ltd. following the manufacturer’s protocol.