Pacific Biosciences of California Inc (PACB)

[Paulieme]

De Novo Assembly
Targeted Sequencing
Base Modification Detection
Isoform Sequencing
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Microbiology
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Home Applications Isoform Sequencing

Isoform Sequencing

Full-length transcript sequencing, no assembly required

RNA sequencing (RNA-Seq) is a broadly used method for studying gene expression. However, short-read sequencing approaches cannot span full-length transcripts, making it difficult to accurately characterize the diverse landscape of isoforms from alternative splicing and transcriptional regulation.

Using the extraordinarily long reads generated by SMRT Sequencing, the Iso-Seq method provides reads that span entire transcript isoforms, from the 5' end to the 3' polyA-tail. It is now possible to directly sequence full-length transcripts ranging up to 10 kb. Generation of accurate, full-length transcript sequences greatly simplifies analysis by eliminating the need for transcript reconstruction to infer isoforms using error-prone assembly of short RNA-Seq reads.

The Iso-Seq method has been applied in a wide variety of organisms to improve annotations in reference genomes, characterize alternatively spliced isoforms in important gene families, and find novel genes in the most comprehensively studied human cell lines. In-depth isoform sequencing can be done across the entire transcriptome or for targeted genes. Understanding the complete representation of a sample's gene isoforms increases the sensitivity and specificity of quantitative functional genomics studies. Isoform sequencing also provides information to efficiently detect or validate novel gene fusions and has been used to determine allele-specific isoform expression.

Benefits:
•Sequence full-length mRNA transcripts, with no assembly required
•Characterize gene-isoform expression across an entire transcriptome, or within targeted genes
•Discover novel genes and gene isoforms even in well characterized samples
•Perform de novo gene annotation, with or without a reference genome
•Complete information about alternatively spliced exons, transcriptional start sites, polyadenylation sites and strand orientation
•Improve quantitation accuracy for functional genomics studies
http://www.pacb.com/applications/isoseq/index.html