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Application note: Optimising read length for human genome sequencing

Literature

Date: 1st July 2021

Introduction

The ability to obtain sequencing reads of any length with nanopore technology, whilst maintaining base modification information, provides a unique opportunity to access the human genome in a way that has not been previously possible. Such comprehensive characterisation of the human genome, from detecting single nucleotide variation (SNV) and nucleotide-level methylation differences to calling large-scale structural variants (SVs) and chromosomal rearrangements, requires careful consideration of the end-to-end experimental workflow — from sample preparation to sequencing and data analysis.

The high-throughput, modular Oxford Nanopore PromethIONTM benchtop sequencer is ideal when it comes to human genome analysis, providing the capacity to run up to 24 (P24) or 48 (P48) flow cells at any time, allowing unprecedented flexibility for users’ experimental requirements.

Objectives

The aim of this Application note is to familiarise users with different sample and library preparation options available for human whole-genome sequencing, based on the combination of Circulomics and Oxford Nanopore Technologies kits and workflows, and the typical sequencing metrics that are produced from each.

With the inclusion of application-based recommendations, this Application note also intends to highlight key experimental considerations at each stage of a genome sequencing workflow.

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