RNA-Seq of Cunninghamia lanceolata:male cone
Ethanol precipitation protocol and CTAB-PBIOZOL reagent was used for the purification of total RNA according to the manual instructions. Total RNA was qualified and quantified by a Nano Drop and Agilent 2100 bioanalyzer (Thermo Fisher Scientific, MA, USA). Oligo(dT)-attached magnetic beads were used to purified mRNA. mRNA were then fragmented into small pieces. First-strand cDNA is generated in First Strand reaction system, and the second-strand cDNA as well. Afterwards, the purified cDNA were connected with adapters. Then, cDNA fragments were amplified by PCR and then purified by Ampure XP Beads. Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. Then the library is sequenced using Illumina HiSeq 4000 platform (BGI-Shenzhen, China) (reads length 151bp).