Quantifying Viral Load via qRT-PCR

Viral load was quantified in capsid gene copy number units using the qRT-PCR method essentially as described previously [14 (link)]. Reactions were performed in duplicate using the SensiFAST SYBR No-ROX One-Step kit (Bioline, Alexandra, NSW, Australia) on a BioRad CFX96/C1000 thermal cycler platform using the primers RHDV-RT2_fw 5’-ACCCAGTACGGCACRGGCTCCCAACCAC-3′ and RHDV-RT2_rv 5’-CTATCTCCATGAAACCAGATGCAAAGGT-3′ [15 (link)].

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Hall R.N., Capucci L., Matthaei M., Esposito S., Kerr P.J., Frese M, & Strive T. (2017). An in vivo system for directed experimental evolution of rabbit haemorrhagic disease virus. PLoS ONE, 12(3), e0173727.

Publication 2017

SensiFAST SYBR No-ROX One-Step kit CFX96/C1000

Capsid Primers

Corresponding Organization : University of Sydney

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Variable analysis

independent variables

QRT-PCR method

dependent variables

Viral load in capsid gene copy number units

control variables

Primer sequences RHDV-RT2_fw and RHDV-RT2_rv
SensiFAST SYBR No-ROX One-Step kit
BioRad CFX96/C1000 thermal cycler platform

controls

Reactions were performed in duplicate

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