Transcriptome analysis of Cynanchum komarovii  under cold stress
A total amount of 3 µg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s recommendations. Briefly, mRNA was purified from total RNA using polyT oligo-attached magnetic beads (Novogene, China). First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase. Second strand cDNA synthesis was subsequently performed using DNA polymerase I and RNase H. In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA) and library quality was assessed on the Agilent Bioanalyzer 2100 system. After cluster generation, four libraries (CK_1, CK_2, CT_1 and CT_2) preparations were sequenced on the Illumina HiseqTM 2000 platform and paired-end reads were generated.