Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

GUS staining and quantitative analysis

JL Jianfeng Lei
PD Peihong Dai
YL Yue Li
WZ Wanqi Zhang
GZ Guantong Zhou
CL Chao Liu
XL Xiaodong Liu
request Request a Protocol
ask Ask a question
Favorite

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.

Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A