Transcriptome and whole-genome resequencing of osmanthus mutant bud of bud mutant branch Osmanthus fragrans
DNase I degradeD double-stranded and single-stranded DNA presenting in RNA samples.Total RNA samples were treated with Ribo-Zero Magnetic Kit to deplete rRNA.RNA molecules were fragmented into small pieces using fragmentation reagent.First-strand cDNA was generated using random primers reverse transcription, followed by a second-strand cDNA synthesis.The synthesized cDNA was subjected to end-repair and then was 3 adenylated. Adapters were ligated to the ends of these 3 adenylated cDNA fragments.Several rounds of PCR amplification with PCR Primer Cocktail and PCR Master Mix are performed to enrich the cDNA fragments. Then the PCR products are purified with Ampure XP Beads.The libraries were assessed quality and quantity in two methods: check the distribution of the fragments size using the Agilent 2100 bioanalyzer, and quantify the library using real-time quantitative PCR (QPCR) (TaqMan Probe).The qualified libraries were amplified on cBot to generate the cluster on the flowcell. And the amplified flowcell will be sequenced pair end on the Illumina System.