RNA sequencing for plant + animal sciences

The PacBio isoform sequence (Iso-Seq) method generates full-length cDNA sequences — from 5’ to the poly-A tail — to confidently characterize the full transcriptome.

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Capture full-length transcripts with no assembly required

The Iso-Seq method employs highly accurate long reads (HiFi reads) to survey transcript diversity, either broadly or in a targeted fashion, providing high-quality genome annotation and gene discovery power.

  • Discover novel genes and informs even in well-characterized model organisms
  • Characterize alternative start and polyadenylation sites as well as exon skipping events
  • Create a reference transcriptome for better RNA-Seq quantification at isoform-level resolution

Workflow: from RNA to full-length transcripts

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Sample + library preparation

Prepare full-length transcripts with flexible sample and library workflows.

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Sequencing

Simplify sample prep workflow and reduce project costs by using the Sequel II and IIe systems.

Scale throughput on Sequel II and IIe systems

  • Use the Sequel II and IIe systems to generate up to 4 million full-length reads per SMRT Cell 8M*
  • Or use the Sequel system to generate up to 500,000* FLNC reads per SMRT Cell 1M

*Read lengths, number of reads, data per SMRT Cell, and other sequencing performance results can vary based on sample quality/type and insert size, etc.

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Data analysis

Output high-quality, full-length transcript sequences; no assembly required.

  • Generate highly accurate long reads (HiFi reads) to characterize whole transcriptomes
  • Use the Iso-Seq analysis in SMRT Link to output high-quality, full-length transcript FASTA sequences, with no assembly required, to characterize transcripts and splice variants
  • Run Iso-Seq analysis with or without a reference genome. Follow up with community tools for different analyses such as SQANTI3 (transcript classification), tappAS (differential expression analysis), and TAMA (annotation).

 

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Application brief

Long-read RNA sequencing

Learn more about these best practices for long-read RNA sequencing

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Spotlight

ISO-SEQ ANALYSIS PROVIDES INSIGHTS INTO FEATS OF PHYSIOLOGY OF HIBERNATING BEARS

Washington State University researchers use Iso-Seq analysis to shed light on the metabolic mystery of brown bears.

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