RT Journal Article
SR Electronic
T1 Rapid High Throughput Whole Genome Sequencing of SARS-CoV-2 by using One-step RT-PCR Amplification with Integrated Microfluidic System and Next-Gen Sequencing
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.11.04.369165
DO 10.1101/2020.11.04.369165
A1 Tao Li
A1 Hye Kyung Chung
A1 Papa K. Pireku
A1 Brett F. Beitzel
A1 Mark A. Sanborn
A1 Cynthia Y. Tang
A1 Richard Hammer
A1 Detlef Ritter
A1 XiuFeng Wan
A1 Irina Maljkovic Berry
A1 Jun Hang
YR 2020
UL http://biorxiv.org/content/early/2020/11/05/2020.11.04.369165.abstract
AB The long-lasting global COVID-19 pandemic demands timely genomic investigation of SARS-CoV-2 viruses. Here we report a simple and efficient workflow for whole genome sequencing utilizing one-step RT-PCR amplification on a microfluidic platform, followed by MiSeq amplicon sequencing. The method uses Fluidigm IFC and instruments to amplify 48 samples with 39 pairs of primers in a single step. Application of this method on RNA samples from both viral isolate and clinical specimens demonstrate robustness and efficiency of this method in obtaining the full genome sequence of SARS-CoV-2.