Confirming Mutant Strain Δ_gidA_ via RT-PCR

To confirm the mutant strain ΔgidA, we performed RT-PCR according to our previously reported methods (Tan et al., 2015 (link)). Briefly, RNA was isolated using SV Total RNA Isolation System (Promega, USA) according to the manufacturer's instructions. In addition, cDNA was synthesized using HiScript Q Select RT SuperMix (Vazyme, China) according to the manufacturer's instructions.
To confirm whether the upstream and downstream genes of gidA are unaffected and functioning normally, we designed the primers of SSU05_2162, gidA, and SSU05_2164 for RT-PCR (Table 2) from the cDNA.

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Gao T., Tan M., Liu W., Zhang C., Zhang T., Zheng L., Zhu J., Li L, & Zhou R. (2016). GidA, a tRNA Modification Enzyme, Contributes to the Growth, and Virulence of Streptococcus suis Serotype 2. Frontiers in Cellular and Infection Microbiology, 6, 44.

Publication 2016

SV Total RNA Isolation System HiScript Q Select RT SuperMix

Cdna Genes Isolation Primers Promega Rt pcr Strain

Corresponding Organization : Center of Hubei Cooperative Innovation for Emissions Trading System

Other organizations : Animal Science Research Institute, Hubei Academy of Agricultural Sciences

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Variable analysis

independent variables

Primers for SSU05_2162, gidA, and SSU05_2164

dependent variables

Confirmation of whether the upstream and downstream genes of gidA are unaffected and functioning normally

control variables

RNA isolation using SV Total RNA Isolation System (Promega, USA)
CDNA synthesis using HiScript Q Select RT SuperMix (Vazyme, China)

controls

Positive control: Not specified
Negative control: Not specified

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