Validation of Suitable Reference Genes for Quantitative Gene Expression Analysis in Panax ginseng

Frontiers in Plant Science
Meizhen Wang, Shanfa Lu

Abstract

Reverse transcription-qPCR (RT-qPCR) has become a popular method for gene expression studies. Its results require data normalization by housekeeping genes. No single gene is proved to be stably expressed under all experimental conditions. Therefore, systematic evaluation of reference genes is necessary. With the aim to identify optimum reference genes for RT-qPCR analysis of gene expression in different tissues of Panax ginseng and the seedlings grown under heat stress, we investigated the expression stability of eight candidate reference genes, including elongation factor 1-beta (EF1-β), elongation factor 1-gamma (EF1-γ), eukaryotic translation initiation factor 3G1 (IF3G1), eukaryotic translation initiation factor 3B (IF3B), actin (ACT), actin11 (ACT11), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and cyclophilin ABH-like protein (CYC), using four widely used computational programs: geNorm, Normfinder, BestKeeper, and the comparative ΔCt method. The results were then integrated using the web-based tool RefFinder. As a result, EF1-γ, IF3G1, and EF1-β were the three most stable genes in different tissues of P. ginseng, while IF3G1, ACT11, and GAPDH were the top three-ranked genes in seedlings treated with heat. Using thre...Continue Reading

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Jul 22, 2014·Journal of Integrative Plant Biology·Meizhen WangShanfa Lu
Nov 8, 2014·Journal of Ginseng Research·Murukarthick JayakodiTae-Jin Yang

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Citations

Sep 21, 2016·International Journal of Molecular Sciences·Lauralie Mangeot-PeterGea Guerriero

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Datasets Mentioned

BETA
KU215664
AAO42312.1

Methods Mentioned

BETA
RNA-seq
PCR
electrophoresis

Software Mentioned

geNorm
Normfinder
BestKeeper
CFX
CFX [UNK]
Primer
coffee
RefFinder
Blastx
Excel

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