Long fragment sequencing applications could help us advance genomics by resolving some of the most challenging regions of the human genome, discerning previously inaccessible structural variants and making sense of the deserts of non-coding material. By offering greater context, the full spectrum of genetic variation could be revealed by utilising longer fragments, giving opportunities for the discovery of novel mechanisms of disease.
DNA size selection can exclude short fragments to accelerate long fragment sequencing research. The process maximises data yields by ensuring those fragments with the most informational content are not blocked from accessing detection centres (i.e. nanopores and/or zero mode waveguides) by shorter DNA fragments.
Whole human genome sequencing use cases are characterised by a need for high data yields to achieve appropriate depth of coverage. Utilisation of Ranger® Technology offers industry-leading precision, scale and automation for the electrophoretic size selection workflow that is crucial for securing the most data from every run.
In addition, Ranger® Technology is helping transform our understanding of species diversity, evolution and agricultural engineering across plant and animal genomic research, where long fragment recovery is critical to ensure construction of high-quality assemblies for complex genomes which feature abundant repeats and polyploidy.
PacBio Compatible Partner
The LightBench®, powered by Ranger® Technology, is a three-in-one instrument offering automated DNA size selection, fragment length analysis and fluorometric quantification. It isolates size ranges of target DNA with industry-leading recovery and precision.
LightBench® excludes smaller fragments, optimising long read inputs to deliver increased mean HiFi read lengths and data yields for the Pacific Biosciences sequencer platforms. To support an automated PacBio library prep workflow, the LightBench® can be integrated into 3rd party liquid handling systems thanks to its SiLA 2-compliant API.
Click to read the press release: Yourgene Health Becomes a PacBio Compatible Partner: Enabling PacBio Customers to Optimise Size Selection for Long-Read Sequencing Libraries
Quality Control and Size Selection for PacBio® Long-read Sequencing using the LightBench® Discover at HudsonAlpha’s CoLab
HudsonAlpha’s CoLab has successfully implemented the LightBench® Discover in their PacBio WGS workflow, using it for precise gel-based size selection and fragment length quality control of all samples. Gel-based size selection on the LightBench® Discover consistently removes DNA fragments <10 kb, increasing mean HiFi read lengths. The fragment length distribution data informs real-time decisions at key stages in the workflow, including assessment of HMW DNA quality, confirmation of DNA shearing, and verification of the final DNA library size and quality. The LightBench® Discover offers a reliable solution for integrated size-selection and quality control in long-read sequencing applications.
PacBio Tech Note
SIZE SELECTION OF HIFI LIBRARIES WITH THE LIGHTBENCH INSTRUMENT FROM YOURGENE HEALTH
In this Tech note, we can see how the improved average HiFi data output of >100Gb achievable on the PacBio Revio system is attributable to LightBench and its next-generation DNA size selection function.
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