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      3. 三代全長轉錄組測序
        Updated:2021/12/22


        Introduction

        The study of full-length transcriptome is an important way to understand the function of organisms. Traditional second-generation sequencing can not directly obtain a single RNA molecule sequences from 5 ˊ To 3 ˊ. The transcriptome research based on Pacbio third-generation sequencing platform can directly read the full-length cDNA of reverse transcription without interruption, effectively obtain all sequences of high-quality single RNA molecule, and accurately identify the transcripts of isoform, homologous gene, superfamily gene or allele expression that cannot be recognized by second-generation sequencing.


        1 Content of bioinformatics analysis

        1.1 Sequencing quality analysis, statistics and evaluation

        1.2 Full length sequence judgment, high quality isoform (qv > 99%)

        1.3 Reference genome alignment; 3. Prediction of new transcripts

        1.4 Quantitative analysis of gene expression and analysis of gene expression level

        1.5 Differential gene expression analysis, differential gene go enrichment analysis and pathway enrichment analysis

        1.6 SNP and indel analysis

        1.7 Transcript structure analysis: variable shear analysis; Fusion gene analysis


        2 Sample requirements

        2.1 total amount > 10 μg RNA

        2.2 Concentration > 50 ng/μL

        2.3 260/280 > 2.0


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