Total Arabidopsis RNA was isolated according to the manufacturer's protocol using an Isolate RNA Minikit (Bioline, NJ). cDNA was synthesized using a high‐capacity DNA reverse transcription kit (Applied Biosystems, CA). RT‐qPCR reactions were done using a 7500 Fast Real‐Time PCR system (Applied Biosystems, CA) as described earlier (Mooney et al., 2019 (link)). Arabidopsis Actin2 (At3g18780) was used as the internal control gene, and all experiments were repeated at least three times as biological replicates, if not otherwise stated. For in planta detection of U‐PEST and U‐SBC expression specific primer pairs were designed that covered eK and PEST or SBC regions, respectively. All primers used are listed in Table S1 . The 2(‐Delta Delta C(T)) method was used to calculate relative gene expression (Livak & Schmittgen, 2001 (link)).
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Arabidopsis RNA Extraction and RT-qPCR Analysis
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Corresponding Organization : Washington State University
Other organizations : Tafila Technical University, Osnabrück University
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Variable analysis
independent variables
- U-PEST expression
- U-SBC expression
- Relative gene expression
- Arabidopsis Actin2 (At3g18780) (internal control gene)
- Positive control: None mentioned
- Negative control: None mentioned
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