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A benchmarking study of SARS-CoV-2 whole-genome sequencing protocols using COVID-19 patient samples

By Tiantian Liu, Zhong Chen, Wanqiu Chen, Xin Chen, Maryam Hosseini Asanjan, Zhaowei Yang, Jing Li, Diana Ho, David Turay, Ciprian Gheorghe, Wendell Jones, Charles Wang

Posted 10 Nov 2020
bioRxiv DOI: 10.1101/2020.11.10.375022

The COVID-19 pandemic is a once-in-a-lifetime event, exceeding mortality rates of the flu pandemics from the 1950's and 1960's. Whole-genome sequencing (WGS) of SARS-CoV-2 plays a critical role in understanding the disease. Performance variation exists across SARS-CoV-2 viral WGS technologies, but there is currently no benchmarking study comparing different WGS sequencing protocols. We compared seven different SARS-CoV-2 WGS library protocols using RNA from patient nasopharyngeal swab samples under two storage conditions. We constructed multiple WGS libraries encompassing three different viral inputs: 1,000,000, 250,000 and 1,000 copies. Libraries were sequenced using two distinct platforms with varying sequencing depths and read lengths. We found large differences in mappability and genome coverage, and variations in sensitivity, reproducibility and precision of single-nucleotide variant calling across different protocols. We ranked the performance of protocols based on six different metrics. Our results indicated that the most appropriate protocol depended on viral input amount and sequencing depth. Our findings offer guidance in choosing appropriate WGS protocols to characterize SARS-CoV-2 and its evolution. ### Competing Interest Statement The authors have declared no competing interest.

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