RNA seq of Cynodon dactylon: second stem internode
The second internodes of the stems at the same growth stage (2 week after tillering) were cut and immediately frozen in liquid nitrogen. Total RNA was extracted using RNAprep pure Plant kit (Tiangen, Beijing, China). RNA quality and quantity were determined by gel electrophoresis and Nanodrop ND1000 spectrophotometer (Nanodrop Technologies, Rockland, ME, USA), respectively. cDNA libraries were prepared using Illumina TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA, USA) according to the manufacturerĄŻs instructions. Briefly, the mRNA was purified from total RNA using magnetic beads attached with poly-T and fragmented into short fragments, which were transcribed into first-strand cDNA using reverse transcriptase and random primers. The second-strand cDNA was then synthesized using DNA polymerase I. A suitable length (150 bp) of cDNA fragments were selected and amplified by PCR to construct the final cDNA libraries. Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) was used to check the quality of the libraries, which were sequenced using Illumina HiSeqTM 2000 (Illumina, USA)