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Full-length Transcriptome Sequencing
Brief

Iso-seq refers to the use of third-generation sequencing technology to sequence complete RNA molecules in organisms (including the full-length sequence from the 5' end to the 3' poly A tail). The RNA molecule does not need to be interrupted, and the complete sequence can be directly obtained by sequencing. More accurate transcript information can be obtained, and more variable splicing sites, new genes and new isomers, and fusion genes can be identified through Iso-seq.

Fields
  • Reveal the transcriptome complexity of species
  • Assisted genome annotation
  • Explore transcript differences under different treatment conditions
  • Structural and functional specificity of transcripts in different time and tissues
  • Explore the pathogenesis of diseases and tumors through variable splicing and fusion genes
Process
1、Third-generation Sequencing - Full-length Transcriptome Sequencing (Iso-seq).png
Parameters
Product TypeSequencing StrategyRecommended Data Amount
Full-length TranscriptomeConstruct a 1-10kb iso-seq library30 Gb subreads; 50 Gb subreads
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Requirements
 Sampling Requirements
Total RNA≥2μg
RNA Concentration≥230 ng/μl
RNA PurityOD260/280: 2.0-2.2
OD260/230: 1.8-2.1
Other RequirementsRIN≥7.2; 28s:18s≥1.0
Do not freeze and thaw repeatedly
Do not get exposed to high temperature (>65°C)
Do not get exposed to high and low pH (<6 or ≥ 9)
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Sample TypeRecommended Sample Size(No Biological Replicates Required)Recommended Site
Plant Tissue≥3-4gTender tissue
Animal Tissue≥1-2gViscera, gills, etc.
Cell≥1x107 cells/library-
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