Automated Library Construction and Analysis for High-Throughput Nanopore Sequencing of SARS-CoV-2.

ABSTRACT:

Background

To support the implementation of high-throughput pipelines suitable for SARS-CoV-2 sequencing and analysis in a clinical laboratory, we developed an automated sample preparation and analysis workflow.

Methods

We used the established ARTIC protocol with approximately 400 bp amplicons sequenced on Oxford Nanopore's MinION. Sequences were analyzed using Nextclade, assigning both a clade and quality score to each sample.

Results

A total of 2179 samples on twenty-five 96-well plates were sequenced. Plates of purified RNA were processed within 12 h, sequencing required up to 24 h, and analysis of each pooled plate required 1 h. The use of samples with known threshold cycle (Ct) values enabled normalization, acted as a quality control check, and revealed a strong

SUBMITTER: Coope RJN 

PROVIDER: S-EPMC9384306 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature