ScienceAsia 48 (2022): 833-838 |doi:
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
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* Corresponding author, E-mail: email@example.com
Received 26 Nov 2021, Accepted 7 Jul 2022