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Research articles

ScienceAsia 48 (2022): 833-838 |doi: 10.2306/scienceasia1513-1874.2022.124


Screening of suitable reference genes for gene expression using quantitative real-time PCR in Gynura bicolor DC.


Hua Xu*, Yemei Fang, Xing Zhang, Chuanna Fan, Xiaoman Sun, Ting Chen

 
ABSTRACT:      Quantitative real-time PCR (qRT-PCR) is a preferred approach for monitoring gene expression levels. Screening of the most stable reference genes is paramount to the accuracy of data generated by qRT-PCR. To date, a systematic exploration of suitable reference genes in different tissues at different developmental stages in G. bicolor has not been performed. Thus, in the present study, a systematic reference gene screening was performed by qRT-PCR on 5 candidate genes in G. bicolor. All 5 reference genes displayed a wide range of Cq values in all samples, indicating that they were expressed variably. Based on the analysis results from the software programs geNorm, NormFinder and BestKeeper, GAPDH was the most stable reference gene in roots and leaves during different developmental stages. ACTIN and GAPDH were selected as the most suitable reference genes in stems. ACTIN was selected as the most suitable gene in all samples, as it was expressed stably in all samples. Overall, this research provides a guideline for the selection of suitable reference genes in qRT-PCR gene expression studies of different tissues at different development stages in G. bicolor.

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* Corresponding author, E-mail: ahxuhua@163.com

Received 26 Nov 2021, Accepted 7 Jul 2022