As opposed to Illumina sequencing, which generates ~150bp reads, Long Read Sequencing produces reads from 1,000 to 1,000,000+ bp. Long Read technology is ideally suited for identifying chromosomal rearrangements, gene fusions, splicing isoforms, and post-transcriptional modifications.
||The 10X Genomics Long Read Genome technology provides long range reads on a genome-wide scale. Combining the 10X Chromium technology with traditional short-read Illumina sequencing, long reads up to 10,000,000+ bp can be reconstructed. Ideal for variant calling, phasing and extensive characterization of genomic structure. DNA extraction from cell culture is available for select cell lines. Flexible bioinformatics approaches are available: FASTQ files, primary Cell Ranger analysis for chromosomal rearrangements, custom secondary analyses|
||Currently offered on a collaborative basis, OICR can perform direct sequencing of either DNA or RNA from samples prepared specifically with an eye to extracting the largest nucleic acids possible. If this interests you, please contact us.|
|Pacific Biosciences Standard libraries||
||The Pacific Biosciences RS II platform provides a reliable means of sequencing long strands of unsheared DNA. This capability allows for sequencing genomes that may not have well characterized reference genomes to align to. It also allows for a more accurate picture when sequencing highly repetitive regions which can be difficult to align to a reference using Illumina short read sequencing. Long read sequencing enables easy detection and characterization of gene fusions and rearrangements.|
|Pacific Biosciences RNA Iso-Seq||
||The isoform sequencing (Iso-Seq) application generates full-length cDNA sequences eliminating the need for transcriptome reconstruction using isoform-inference algorithms. The Iso-Seq method generates accurate information about alternatively spliced exons and transcriptional start sites. Full-length cDNA sequences can be used by themselves or in conjunction with traditional Illumina short read sequencing to infer chromosomal rearrangements that result in fusion transcripts.|
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This technical note describes considerations and best practices for 10X Linked-Reads experiments, as well as examples of quality control metrics used to evaluate run performance and detect possible outliers within projects. Includes sequencing guidelines for various 10X Linked-Reads applications.