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Full-length Transcriptome Sequencing

Relying on advanced sequencing systems and cutting-edge technologies, Creative Proteomics provides global customers with full-length transcriptome sequencing and analysis services, aiming to help you improve the genome annotation of species, perform more accurate quantitative analysis and alternative splicing analysis.


Full-length Transcriptome Sequencing

The full-length transcriptome is based on single-molecule real-time sequencing technology, which can directly obtain full-length transcript information without assembly. Its ultra-long read length ensures the accuracy of transcriptome sequencing results to a greater extent. At the same time, it complements the gene annotation results of the annotated genomes, discovers new genes and transcripts, identifies alternative splicing, gene fusion phenomena, alternative polyadenylation sites (APA), etc., and improves gene Expression quantification. Therefore, it overcomes the problems of short splicing and incomplete information of transcripts without reference genome species, and realizes the study of structural variations such as alternative splicing and fusion genes in species with reference genome.

Advantages of full-length transcriptome sequencing and analysis:

  • No assembly required: Ultra-long reads do not require assembly, and the sequence information of full-length transcripts can be accurately obtained.
  • High throughput: Low cost of sequencing and short cycle time.

Applications of full-length transcriptome sequencing and analysis:

  • Refine the genome annotation of species.
  • More accurate quantitative analysis.
  • Changes in variable splicing events induced by different experimental treatments.
  • Research on post-transcriptional regulatory mechanisms, such as variable polyadenylation.
  • Fusion gene studies in carcinogenesis.
  • Obtain more complete reference coding sequences for species without reference genomes.

Advantages of Our Full-length Transcriptome Sequencing and Analysis Services

  • Comprehensive information analysis content: Creative Proteomics follows the frontiers of scientific research in real time, and constantly upgrades the information analysis content.
  • Personalized analysis: With rich experience in personalized analysis, more suitable analysis software can be selected according to project needs, just to ensure more accurate results.
  • Rich experience: Thousands of full-length transcriptome sequencing projects have been completed, the platform is running well, and rich experience in dealing with problems in experiments, sequencing, and bioinformatics analysis.

Service Workflow

For full-length transcriptome sequencing of experimental samples, we followed the basic workflow shown below. At the same time, Creative Proteomics has developed a variety of novel sample preparation techniques, and we will adjust the experimental methods and optimize the experimental conditions according to the different properties of the samples.

Service Workflow of Full-length Transcriptome Sequencing -Creative Proteomics

Sample Requirements

  • Total RNA: ≥ 5 μg, concentration ≥ 300 ng/μL, RIN ≥ 8.0, OD260/280 ≥ 1.8, OD260/230 ≥ 1.8; The baseline is flat and the 5S peak is normal.
  • Tissue: fresh animal tissue dry weight = 50-200 mg, fresh plant tissue dry weight = 500-1000 mg.
  • Whole blood: mammalian whole blood = 2-5 mL, non-mammalian whole blood = 0.5-2 mL
  • Freshly cultured cells: ≥ 5×107
  • Bacteria: ≥ 2×107
  • Algae: ≥ 2×107, or ≥ 1 g

Bioinformatics Analysis

Type Analyze content
Have a reference genome

Standard bioinformatics analysis

  • Data quality control, transcript clustering and calibration
  • Alignment with reference genome
  • New transcript prediction
  • New transcript functional annotations (Nr, GO, KEGG, SwissProt)
  • Quantitative analysis as a reference sequence
  • Gene structure analysis
    • lncRNA analysis
    • Alternative splicing analysis
    • Variable polyadenylation analysis
    • Fusion gene analysis
    • SNP/InDel analysis
    • Open reading frame (ORF) analysis
    • Transcription factor (TF) analysis
    • Gene structure optimization

Customized information analysis

  • Use the NGS data to correct the third - generation sequencing data
  • Gene quantification and differential expression analysis
  • Multi-omics integrated analysis (eg methylation, proteome, miRNA)
No reference genome

Standard bioinformatics analysis

  • Data quality control, transcript clustering and calibration
  • Gene functional annotation (Nr, GO, KEGG, SwissProt)
  • High-level functional annotation of genes
    • Predicted coding protein box (CDS)
    • Transcription factor analysis (TF)
    • R gene analysis (plant)
    • Protein domain analysis (Pfam, SMART)
    • TMHMM transmembrane helix structure prediction
    • SignalP signal peptide structure prediction
    • Protein O-GlcNAc glycosylation sites prediction (mammalian)
    • ProP furin cleavage site prediction (eukaryotes)
  • Structural analysis
  • SSR analysis
  • lncRNA analysis
  • Alternative splicing Analysis

Customized information analysis

  • Use the NGS data to correct the third - generation sequencing data
  • Gene quantification and differential expression analysis
  • Multi-omics association analysis (eg methylation, proteome, miRNA)

If you are interested in full-length transcriptome sequencing, or you want to open up new research, you need to find a professional full-length transcriptome sequencing and analysis company for cooperation. Don't hesitate to get in touch with us today, Creative Proteomics can be trusted.

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