Transcriptome analysis of Chinese bayberry (Myrica rubra Sieb. et Zucc.) fruit treated with heat and 1-MCP
The total RNA were extracted from the Chinese bayberry fruits at different stages, CK0, CK6, HM6 with two biological replicates (CK0_1, CK0-2, CK6_1, CK6_2, HM6_1 and HM6_2 respectively) according to physiological analysis results having two replicates for each. The modified CTAB method was used to extract total RNA and then to construct the transcriptome libraries (Qian et al. 2014). Different techniques like NanoDrop 2000 (Thermo Fisher, CA, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA) were used to evaluate RNA purity and integrity. DNase were used to remove the contaminating genomic DNA. After the quality control procedure, mRNA enriched form total RNA using Oligo (dt) beads was fragmented randomly in the fragmentation buffer, then the cDNA was synthesized using a random hexamers and reverse transcriptase. The final cDNA library was ready after a round of purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR enrichment. All libraries were sequenced by Illumina HiSeqTM 4000 with a paired-end run. The original raw data from Illumina HiSeqTM 4000 were transformed to Sequenced Reads by base calling. Remove reads containing adaptors or low-quality reads from raw data to get clean data used in the following analysis (1GENE.biotech.co.ltd provided technical support for RNA-seq and data analysis).