Single Molecule, Real-Time (SMRT) Sequencing technology delivers exceptional read lengths without compromising throughput or accuracy.
Retrieving HiFi sequencing reads up to 30kb, long read sequencing overcomes the challenges that short-reads have in difficult regions such as repetitive elements and large structural variations.
Long Read Sequencing
Genomic, transcriptomic and epigenetic characterization with high precision
Why choose long read sequencing?
Discuss your project with me
Main advantages
Next generation sequencing (NGS) technologies typically make use of generating 75-300bp long reads, which are then either de-novo assembled in a genome or mapped to a reference genome. Complex regularity elements or highly repetitive sequences can make this mapping harder than for uniquely variable ones. By utilizing SMRT technology of the PacBio Sequel II system, we are able to easily generate highly accurate (median consensus accuracy of >99.9%) long reads up to 30 kb. With reads tens of kilobases in length you can readily assemble complete previously uncharacterized genomes, sequence full-length transcripts as well as genes or regions of interest ranging in size from hundreds base pairs to 30 kb.
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Easier assembly of complete genomes
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Sequencing full-length RNA transcripts
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Direct detection of epigenetic modifications
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Haplotype phasing information
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Base-level resolution with >99.9% single-molecule read accuracy
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Unbiased coverage of GC rich regions such as promoters
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No PCR amplification, prevention of errors
Additionally, calling and detection of variants benefit from long-read sequencing thanks to low sequencing-context bias, and accurate mapping of reads. Single Molecule, Real-Time (SMRT) Sequencing is a core technology and a powerful tool for analyzing variable number tandem repeats, or for repeat occurrence with disease significance, such as in Huntington’s disease.
Service workflow
1. Initial meeting
When you consider using one of our services, our scientific team will help choosing the optimal project design. Each service can be customized by adding options. You will receive an overview of project budget and deliverables, so you can make an informed choice. Once you return your Purchase Order form, your project manager will contact you to discuss the next steps.
2. Sample delivery
Sample specifications for long read sequencing are:
Purified DNA
- Quality: High molecular weight DNA (HMW)
- Validated input/ Minimum volume: application dependent (the scientific team will inform you accordingly based on the type of project)
3. Sample entry QC
After sample arrival, Entry-QC is performed to determinate the sample quality and quantity.
A report with QC results will be provided and results and progression of the project can be discussed.
4. Library prep/QC and sequencing
The library resulting from sample preparation goes into a library QC step to ensure sufficient material per sample and the right library size. Sequencing on PacBio Sequel II:
- Data generated from 2-4 circular consensus sequencing reads (CCS)
- Yield: Up to 20Gb CCS data (application depended)
5. Data QC
The raw data that is generated by the sequencer must be divided to obtain one data file per sample. This is called demultiplexing.
Your project manager checks the data quality by analyzing the quality metrics of the run. Final inspection of the data takes place and the data is either transferred on a hard disk or it is uploaded to the GS customer portal.
6. Reporting
You receive your Project Report together with your data. It contains an overview of the experimental methods used, sample specifications, and a summary of all technical information for publication. Deliverables:
- Raw data files via secure e-transfer or shipment on storage medium
- Quality score Q30 of ≥80% for PE150 reads
- Optional data analysis with comprehensive report
Data analysis
GenomeScan’s bioinformatics experts apply optimized pipelines for preprocessing, assemblies, variant calling and disease annotations to analyze your dataset. With our customizable workflows, we help you receive reliable and publication-ready results.
Bioinformatic pipelines can also be tailored according to your research or clinical needs allowing more in-depth mining of your dataset.
How it works
Service specifications
We have summarized key information about our long-read sequencing service into a service specification sheet.
Let's get the conversation started for your next NGS project
Please either fill in this form or email us directly at info@genomescan.nl
and we will get in touch with you to discuss your requirements
