Full-length transcriptome is an important way to understand the function of organisms. Traditional second-generation transcriptome sequencing can not directly obtain a single RNA molecule composed of 5 ˊ To 3 ˊ All sequences of. 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; Full length sequence judgment, high quality isoform (qv > 99%);
1.2 Reference genome alignment;
1.3 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
Total amount > 4 μg RNA
Concentration > 50 ng / μL
260/280 > 2.0
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