Application Updates: Introducing Iso-Seq Express for Faster RNA Sequencing
Wednesday, June 26, 2019
Seeking to sequence and characterize entire transcriptomes in one go? Our new Iso-Seq protocol and reverse-transcriptase PCR kit makes it easier, speedier and cheaper.
Run on the new Sequel II System, the completely revamped Iso-Seq Express workflow achieves whole transcriptome characterization from a single SMRT Cell 8M delivering up to 400 Gb, and at a third of the cost, or less. Yield has also increased on the Sequel System, with 3.0 sequencing chemistry typically delivering up to 30 Gb per SMRT Cell 1M for our RNA sequencing application.
The new protocol requires three times less RNA input (300 ng) and minimizes handling-induced cDNA damage. Preparation is also simplified, allowing you to go from total RNA to SMRTbell library in one day. And no need for size selection: the ProNex bead system (used in place of AMPure PB beads) allows you to tune the size preference of full-length transcripts.
Early access users have praised the new protocol for its ease, output and speed. “Never got this much data and long polymerase read length,” noted one customer. “cDNA length is also way better than any previous data.”
The new Iso-Seq Express also supports multiplexing up to 12 samples for additional savings. This will be particularly useful for researchers interested in capturing gene diversity expressed from different tissues, timepoints, or experimental conditions.
In addition, our improved Iso-Seq workflow in SMRTLink 7.0 allows you to analyze your data and identify full-length transcripts with a faster turnaround time.
Identify, Annotate, Characterize
For plant and animal researchers who need high-quality gene models, full-length transcript sequencing is a key tool for annotating reference genomes or providing a reference transcriptome in the absence of a genome.
For human genetics researchers who are trying to identify disease-causing variants in rare and Mendelian disorders, the improvements allow unambiguous characterization of complex alternative isoforms.
Transcriptome sequencing has also proven important for understanding the functional consequences of cancer genome mutations, including structural rearrangements. The genomes of the COLO 829 melanoma and matched normal peripheral blood cell lines were recently sequenced to 50-fold coverage and paired long-read transcriptome data was generated using the updated Iso-Seq Express method. The results were shared at the AACR 2019 Annual Meeting and the complete data set is now available for download.