In Vitro Transcription of Model vRNAs

A 360-nucleotide model vRNA was transcribed in vitro from a cDNA containing the T7 promoter and the noncoding region of the NS gene of WSN (H1N1), VN/1180 (H5N1), or AH/1 (H7N9) virus by using the RiboMax Large Scale RNA Production Systems (Promega) as described previously [34 (link)]. The vRNAs transcribed in vitro were concentrated and purified by using a Spin Column RNA Cleanup & Concentration Kit (Sangon Biotech) and quantified by using NanoDrop (ThermoFisher).

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Li J., Liang L., Jiang L., Wang Q., Wen X., Zhao Y., Cui P., Zhang Y., Wang G., Li Q., Deng G., Shi J., Tian G., Zeng X., Jiang Y., Liu L., Chen H, & Li C. (2021). Viral RNA-binding ability conferred by SUMOylation at PB1 K612 of influenza A virus is essential for viral pathogenesis and transmission. PLoS Pathogens, 17(2), e1009336.

Publication 2021

RiboMax Large Scale RNA Production Systems Spin Column RNA Cleanup & Concentration Kit NanoDrop

Cdna Gene H5n1 H7n9 virus Nucleotide Promega

Corresponding Organization : Harbin Veterinary Research Institute

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Variable analysis

independent variables

Virus type (WSN (H1N1), VN/1180 (H5N1), or AH/1 (H7N9))

dependent variables

Concentration and purity of in vitro transcribed vRNA

control variables

T7 promoter sequence
Noncoding region of the NS gene
RiboMax Large Scale RNA Production Systems (Promega) protocol
Spin Column RNA Cleanup & Concentration Kit (Sangon Biotech)
NanoDrop (ThermoFisher) for quantification

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