title: Transcriptome sequencing
design: There are two main ways to obtain mRNA: one is to use the structural features of most eukaryotic mRNAs with polyA tails, and the mRNAs with polyA tails are enriched by Oligo(dT) magnetic beads; the other is to remove them from total RNA. ribosomal RNA to obtain mRNA. Fragmentation buffer was then added to break the RNA into short fragments. The short fragment RNA was used as a template to synthesize the first-strand cDNA with random hexamers. Then buffer, dNTPs (dUTP, dATP, dGTP and dCTP) were added. The double-stranded cDNA was synthesized with DNA polymerase I, and the double-stranded cDNA was purified by AMPure XP beads. The purified double-stranded cDNA is then subjected to end repair, A tail is added, and the sequencing adapter is connected, and then AMPure XP beads are used for fragment size selection, and finally PCR enrichment is performed to obtain the final cDNA library.Different libraries are pooled according to the target data volume, and sequenced on the Illumina Novaseq platform