NODETranscriptomicshua liRNA-Seqhttps://www.biosino.org/node/experiment/detail/OEX00019944Illumina NovaSeq 6000PairedPCRN11['cell Line']['Homo sapiens']Total RNA was extracted from the tissue according to the instructions (Invitrogen) using TRIzol ®reagents, and genomic DNA was removed using DNase I (TaKara). Then the mass of RNA was determined by 2100 biological analyzer (Agilent), and the quantity was determined by ND-2000 (NanoDrop Technologies). High quality RNA samples (OD260/280=1.8~2.0,OD260/230 ≥ 2.0 ng / μ l, concentration ≥ 100 ng / μ l, total amount ≥ 2 μ g) were used for subsequent database construction. A double-terminal library was constructed using ABclonal mRNA-seq-Lib preparation kit (ABclonal,China), and mRNA was purified from 1 μ g total RNA using oligo (dT) magnetic beads, and then cleaved with divalent cations in the first chain synthesis buffer of ABclonal. Then, the first strand cDNA was synthesized by random hexamer primers and reverse transcriptase (RNAseH) using mRNA fragment as template, and then the second strand cDNA was synthesized by DNA polymerase I, RNAseH, buffer and dNTPs. The synthesized double-stranded cDNA fragments were acidified by polyadenylate and ligated to prepare a double-terminal library. CDNA was then purified by AMPure XP system (Beckman Coulter,Beverly,MA,USA) and amplified by PCA. PCR amplification was performed with adapter-linked cDNA and adapter primers. Finally, Illumina Novaseq 6000 instrument was used for sequencing. The Clean Reads was sequenced with the designated genome using HISAT2 software to obtain its location information on the reference genome. Then the FPKM value (expected number of Fragments Per Kilobase of transcript sequence perMillions base pairs sequenced) of each gene expression in each sample was calculated by featureCounts software. Then, the differential expression of genes was analyzed by Deseq2 software. The differential genes were screened according to the criteria of difference multiple > 2 and p-value < 0.05.Results: 1.NODEfalseRNA-SEQRNA sequencing library preparation and analysis2022-07-082022-07-08OEX00019944OEP00003511