ppat.1009402.s002.tif (1.49 MB)

Murine norovirus (MNoV) hybridization capture yields improved genomic sequencing efficiency, dependent upon initial viral levels.

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posted on 2021-03-11, 18:54 authored by Forrest C. Walker, Ebrahim Hassan, Stefan T. Peterson, Rachel Rodgers, Lawrence A. Schriefer, Cassandra E. Thompson, Yuhao Li, Gowri Kalugotla, Carla Blum-Johnston, Dylan Lawrence, Broc T. McCune, Vincent R. Graziano, Larissa Lushniak, Sanghyun Lee, Alexa N. Roth, Stephanie M. Karst, Timothy J. Nice, Jonathan J. Miner, Craig B. Wilen, Megan T. Baldridge

(A) Schematic of enrichment protocol, in which cDNA from stool RNA undergoes Nextera tagmentation, followed by hybridization capture using biotinylated MNoV-specific probes and streptavidin beads (yellow hexagons) for pulldown prior to Illumina sequencing. (B) Stool samples collected from various mice infected PO with CR6 underwent this protocol, then pre- and post-enrichment samples were sequenced to assess efficacy of enrichment. (C) For stool samples from this study (Figs 1D and S1C) that were deep-sequenced after enrichment, the percentage of reads mapping to MNoV were compared to viral levels detected in samples by qPCR. Linear regression analysis for post-enrichment samples (B) and samples for this study (C) was performed using GraphPad Prism 7 software, with the P-value reporting if the slope is significantly non-zero.

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