Chinese fir cambium transcriptome
Illumina HiSeqTM 2000 beads with oligo(dT) were used to isolate poly(A) mRNA from total RNA. Fragmentation buffer was added to create short mRNA fragments (200–700 bp), which were used as templates with random hexamer (N6) primers (Illumina) to synthesize the first-strand cDNA. The second-strand cDNA was synthesized using buffer, dNTPs, RNase H, and DNA polymerase I. Short fragments were purified with a QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and addition of poly(A). The short fragments were then connected with sequencing adapters. For PCR amplification, suitable fragments were selected as templates, according to the results of agarose gel electrophoresis. Finally, the transcriptome library was paired-end sequenced through a Illumina HiSeqTM 2000 platform at the Beijing Genomics Institute (BGI)-Shenzhen (Shenzhen, China) following the manufacturer’s instructions.