Iso-Seq / PacBio Transcriptome Sequencing
“Isoform Sequencing” (Iso-seq) developed by Pacific Biosciences (PacBio), is based on long-read sequencing technology and allows researchers to identify new isoforms with extraordinary precision. Contact us today with any questions or for a no obligation quote.
Isoform Sequencing (Iso-seq) developed by Pacific Biosciences (PacBio), is based on long-read sequencing technology. The unique long-read sequencing feature allows this method to identify new isoforms with extraordinary precision. The Iso-Seq application generates full-length cDNA sequences — from the 5’ end of transcripts to the poly-A tail — eliminating the need for transcriptome reconstruction using isoform-inference algorithms. The Iso-Seq method provides accurate information about alternatively spliced exons and transcriptional start sites. It also reveals information about poly-adenylation sites for transcripts across the full complement of isoforms within targeted genes or the entire transcriptome.
RNA sequencing has become the most frequently used method for the majority of researchers conducting gene expression profiling. However, it is difficult to obtain a complete picture of the transcriptome because short reads cannot accurately assemble complex transcripts.
- Absolute Quantification with UMI Technology
- Greater Transcripts Detection with Multi-throughput Iso-Seq
- Rapid turnaround times and expert support
- Library preparation – Standard Iso-Seq library / Multi- throughput Iso-Seq library/polyA Iso-Seq library
- 20Gb sequencing data per sample is recommended
Recommended: Mass ≥ 3 µg – Concentration ≥ 285ng/µL – RIN ≥7.5 – 28S/18S(23S/16S) ≥1.2
Required: 1.5µg ≤ m < 3 µg – RIN ≥7.5 – 28S/18S(23S/16S) ≥1.2
- Remove the low-quality reads and short reads;
- Identify the full-length, non-chimeric transcripts and non-full-length, non-chimeric transcripts
- Build similarity graph using BLASR, get cluster consensus
- Polish the consensus sequences and get high quality full-length, non-chimeric transcripts
- Merge consensus sequences of all libraries and remove redundancy
- Annotation of the full-length non-chimeric transcripts (Nr、Nt Swissprot、KEGG、GO、COG and Interpro)
- CDS prediction
- SSR prediction
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PacBio Revio Platform
BGI Genomics’ Iso-Seq/PacBio transcriptome sequencing services are executed on PacBio’s Revio platform. The Revio Sequencing System has been recognized for its ability to generate longer reads with greater accuracy and throughput, at a significantly lower cost.
