Processing frozen archival human DNA samples for large-scale SQK-LSK114 Oxford Nanopore long-read DNA sequencing SOP v1

June 2024

Authors:
Alicia Wenghöfer, Kimberly Paquette, Laksh Malik, Breeana Baker, Cedric Kouam, Kimberley J Billingsley

Abstract:
“As part of the GP2 monogenic network we will generate long-read sequencing data to better understand the genetic architecture of Parkinson’s disease. To generate this large-scale Nanopore data we have developed a protocol for processing and long-read sequencing frozen human DNA samples, ideally targeting an N50 of ~30kb and ~30X coverage. However, with archival human DNA samples, we usually see a drop in DNA quality in terms of DNA length. Therefore, this protocol is focused on attempting to achieve the highest N50 and coverage possible from the input available.“

Sage Science Products:
BluePippin with High-Pass Plus gel cassettes, >10kb High-Pass protocol.

Author Affiliations:
Center for Alzheimer’s and Related Dementias, National Institute on Aging, Bethesda, Maryland
Institute of Neurogenetics, University Hospital Schleswig-Holstein, Luebeck, Germany
Laboratory of Neurogenetics, National Institute on Aging, Bethesda, Maryland

Protocols.io
DOI: 10.17504/protocols.io.5jyl82morl2w/v1

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