Evaluation of suitable reference genes for qRT-PCR gene expression normalization in reproductive, vegetative tissues and during fruit development in oil palm

Yeap, Wan-Chin; Loo, Jia Mayne; Wong, Yick Ching; Kulaveerasingam, Harikrishna

doi:10.1007/s11240-013-0382-3

Cited by 59 publications

(54 citation statements)

References 37 publications

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“…The formula of 2 -4Ct was applied to calculate the relative expression levels of housekeeping genes from the raw C q value (Schmittgen and Livak 2008;Yeap et al 2014). M value and V value can be automatically calculated to For all the sample pools, EF1a and GAPDH were the most stable reference genes, with the lowest M value of 0.618.…”

Section: Amplification Efficiency and C Q Variationmentioning

confidence: 99%

Reference gene selection in Artemisia annua L., a plant species producing anti-malarial artemisinin

Liu

Zhao

Wang

et al. 2014

Plant Cell Tiss Organ Cult

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The selection and validation of reference genes are essential for gene expression studies by real-time quantitative PCR. The genetic map of Artemisia annua L., a Chinese medicinal plant species producing anti-malarial artemisinin, has been reported. However, few reference genes of A. annua have been estimated for real-time quantitative PCR until now. In this study, ten putative housekeeping genes, including ACT, UBQ, TUB, 18S rRNA, EF1a, CYP, RPL13D, TUA, RPII and GAPDH, were chosen for identifying expression stability using geNorm and NormFinder software tools in 11 different sample pools, containing those from different plant organs and from plants treated with phytohormones and abiotic stresses. As

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Section: Amplification Efficiency and C Q Variationmentioning

confidence: 99%

Reference gene selection in Artemisia annua L., a plant species producing anti-malarial artemisinin

Liu

Zhao

Wang

et al. 2014

Plant Cell Tiss Organ Cult

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“…Therefore, the stability of reference genes must be validated before their use (Guenin et al 2009). Researchers have validated the stability of candidate reference genes in various species such as Arabidopsis (Remans et al 2008), rice (Jain 2009), wheat (Paolacci et al 2009), potato (Nicot et al 2005, cotton (Wang et al 2013), soybean (Hu et al 2009), oil palm (Yeap et al 2014), and Chinese cabbage .…”

Section: Introductionmentioning

confidence: 99%

“…Genotype, organ, developmental stage, and abiotic stress all affect the stability of housekeeping genes expression Yeap et al 2014). Garlic abiotic stresses include wounding, salt, drought, oxidation, cold, and heat stress.…”

Section: Introductionmentioning

confidence: 99%

Selection and validation of garlic reference genes for quantitative real-time PCR normalization

Liu

Jiang

2015

Plant Cell Tiss Organ Cult

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A stable reference gene is a prerequisite to obtaining reliable quantitative real-time PCR (qPCR) analysis results. Despite the global distribution and nutritional, medicinal and economic value of garlic, reference genes for qPCR have not been reported. Eight garlic housekeeping genes, 18S ribosomal RNA (18S), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor (EF-1a), the a-tubulin (TUA) and b-tubulin (TUB), polyubiquitin (UBQ), and cyclophilin (CYP), were chosen as candidate reference genes. Their stability was evaluated by GeNorm, NormFinder, BestKeeper, the comparative DCt method, and RefFinder. The results demonstrated that TUA, GAPDH, UBQ, CYP, and ACT are the most suitable of the eight reference genes for different garlic genotypes, organs, developmental stages, abiotic stresses and media treatments, respectively. To further validate the stability of these reference genes, the expression levels of the superoxide dismutase gene (SOD) were analyzed. The relative quantification of SOD varied according to the stability of the reference gene, thus highlighting the importance of using suitable reference genes in qPCR. Our results also provide a molecular biological basis for studying the mechanisms of development and stress tolerance in garlic.

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“…The transcripts ID and primers designed for RT-qPCR are listed in Table 1. The PCR parameters were 95°C for 10 min followed by 45 cycles of 95°C for 15 s and 60°C for 30 s. Two endogenous controls transcripts, SLU7 and GRAS were used in parallel for normalisation (Yeap et al, 2014). The relative transcript level was calculated based on a comparative C T method using two reference genes for normalisation.…”

Section: Rt-qpcr Analysismentioning

confidence: 99%

Validation of Differential Gene Expression of Transcriptome Assembly via Nanostring® Technologies Analysis Platform

Othman¹

2018

JOPR

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Evaluation of suitable reference genes for qRT-PCR gene expression normalization in reproductive, vegetative tissues and during fruit development in oil palm

Cited by 59 publications

References 37 publications

Reference gene selection in Artemisia annua L., a plant species producing anti-malarial artemisinin

Reference gene selection in Artemisia annua L., a plant species producing anti-malarial artemisinin

Selection and validation of garlic reference genes for quantitative real-time PCR normalization

Validation of Differential Gene Expression of Transcriptome Assembly via Nanostring® Technologies Analysis Platform

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