Novaseq 6000 mgiseq t7 instrument

The tissues (root, leaf, flower, stem, and fibrous root) of D. asperoides were used for illumina library preparation and sequencing. In brief, the total RNA was extracted from each tissue using TRIzol^® Reagent (Magen). Each total RNA sample was then used for NGS analysis, while equivalent amounts of RNA from roots, leaves, flowers, stems, and fibrous roots were mixed for SMRT analysis.
The first-strand cDNAs were synthesized with random hexamer primers and Reverse Transcriptase (RNase H) using mRNA fragments as templates, followed by second-strand cDNA synthesis using DNA polymerase I, RNAseH, buffer, and dNTPs. Adaptor-ligated cDNA was used for PCR amplification. PCR products were purified (AMPure XP system) and library quality was assessed on an Agilent Bioanalyzer 4150 system. Finally, sequencing was performed with an Illumina Novaseq 6000/MGISEQ-T7 instrument. Raw data obtained from the transcriptome sequencing by removing the adapter sequence and filtering out low-quality reads to gain high-quality clean reads was used for subsequent analysis. Clean data were used to do de novo assembly with Trinity. The assembled transcriptome sequences were compared with five databases (NR, SwissProt, Pfam, GO and KEGG databases) to obtain the annotation information in each database.