Quantitative PCR Analysis of Bt Resistance

The detailed procedure used for real-time quantitative PCR (qPCR) analysis to detect gene expression has been described elsewhere [66 (link)]. Gene-specific primers of MAPK cascade genes were selected in this study (S3 Table), while qPCR primers of Bt resistance-related genes were obtained from our previous studies [30 (link)–33 (link)]. The qPCR reactions were performed with 2.5 × SYBR Green MasterMix Kit (TIANGEN) following the manufacturer’s instructions in the QuantStudio 3 Real-Time PCR System (Applied Biosystems). Four technical repeats and three biological replicates were conducted for each treatment. Relative expression levels of target genes were calculated using the 2^-ΔΔCt method and normalized to the ribosomal protein L32 (RPL32) gene (GenBank accession no. AB180441).

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Guo Z., Kang S., Wu Q., Wang S., Crickmore N., Zhou X., Bravo A., Soberón M, & Zhang Y. (2021). The regulation landscape of MAPK signaling cascade for thwarting Bacillus thuringiensis infection in an insect host. PLoS Pathogens, 17(9), e1009917.

Publication 2021

SYBR Green MasterMix Kit QuantStudio 3 Real-Time PCR System

Biological Expression genes Gene Mapk cascade Primers Quantitative real time pcr analysis Ribosomal protein l32 Sybr green

Corresponding Organization : Chinese Academy of Agricultural Sciences

Other organizations : University of Sussex, University of Kentucky, Universidad Nacional Autónoma de México

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Variable analysis

independent variables

Gene-specific primers of MAPK cascade genes were selected in this study

dependent variables

Relative expression levels of target genes

control variables

The detailed procedure used for real-time quantitative PCR (qPCR) analysis to detect gene expression has been described elsewhere [66 (link)]
QPCR primers of Bt resistance-related genes were obtained from our previous studies [30 (link)–33 (link)]
The qPCR reactions were performed with 2.5 × SYBR Green MasterMix Kit (TIANGEN) following the manufacturer's instructions in the QuantStudio 3 Real-Time PCR System (Applied Biosystems)
Four technical repeats and three biological replicates were conducted for each treatment
Relative expression levels of target genes were normalized to the ribosomal protein L32 (RPL32) gene (GenBank accession no. AB180441)

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