GUS staining and quantitative analysis
This protocol is extracted from research article:
Heritable gene editing using FT mobile guide RNAs and DNA viruses
Plant Methods, Feb 17, 2021; DOI: 10.1186/s13007-021-00719-4

The upper systemic leaves without injection were GUS stained after infiltration according to a previously described protocol [26]. Briefly, to analyze GUS gene expression, RNA was isolated from the upper systemic leaves of plant inoculated CLCrV-A and CLCrV-B empty vector, GUS-sgRNA and Cas9-OE A. thaliana, and reversely transcribed into cDNA. qRT-PCR was performed amplifying the 168 bp GUS gene. After the reaction was completed, relative gene expression levels were calculated using the 2−∆∆Ct method [27] with A. thaliana Actin2 gene as the reference gene. Primers used for PCR amplification are listed in Additional file 1.

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