Identification of suitable reference genes for gene expression studies by qRT-PCR in the blister beetle Mylabris cichorii

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for qRT-PCR assays. This study identifies suitable reference genes for local chilli, Capsicum annuum var MC11 under incident of Cucumber mosaic virus infection. Six candidate genes actin, tub, EF1α, GAPDH, TEF1α and 18SrRNA and three validated Capsicum reference genes UBI-3 ref, β-tub ref and gapdhref were tested against five chilli plant parts stem, shoot, leave, flower and root. The PCR/qRT-PCR results demonstrate only five candidate references genes actin, EF1α, GAPDH, 18SrRNA, and TEF1α that show specific single band of amplicon, without primer dimers and at the targeted sizes. Through qRT-PCR, GAPDH gives single peak in dissociation curve in all plant parts used further fulfilling the characteristic of reference genes.Previous work on validation of reference genes in pepper shows that only UBI-3 suits to C. annuum var MC11 infected CMV, thus we suggest that GAPDH has a potential to be a validated reference gene for C. annuum var MC11 and can be used together UBI-3 for the purpose of data normalization.

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Identification and Validation of Reference Genes for qRT-PCR Studies of Gene Expression inDioscorea opposita

BioMed Research International ◽

10.1155/2016/3089584 ◽

2016 ◽

Vol 2016 ◽

pp. 1-13 ◽

Cited By ~ 7

Author(s):

Xiting Zhao ◽

Xiaoli Zhang ◽

Xiaobo Guo ◽

Shujie Li ◽

Linlin Han ◽

...

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Developmental Stages ◽

Chain Reaction ◽

Qrt Pcr ◽

Expression Studies ◽

Tuber Crop ◽

Polymerase Chain ◽

Gene Expression Studies ◽

Suitable Reference

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for obtaining reliable results for qRT-PCR assays. This study evaluated potential reference genes of Chinese yam (Dioscorea oppositaThunb.), which is an important tuber crop and medicinal plant in East Asia. The expression of ten candidate reference genes across 20 samples from different organs and development stages was assessed. We identified the most stable genes for qRT-PCR studies using combined samples from different organs. Our results also suggest that different suitable reference genes or combinations of reference genes for normalization should be applied according to different organs and developmental stages. To validate the suitability of the reference genes, we evaluated the relative expression ofPE2.1andPE53, which are two genes that may be associated with microtuber formation. Our results provide the foundation for reference gene(s) selection inD. oppositaand will contribute toward more accurate gene analysis studies of the genusDioscorea.

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Selection of Suitable Reference Genes for qRT-PCR Gene Expression Studies in Rice

Methods in Molecular Biology - Rice Genome Engineering and Gene Editing ◽

10.1007/978-1-0716-1068-8_20 ◽

2021 ◽

pp. 293-312

Author(s):

Meng Wang ◽

Navreet K. Bhullar

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Qrt Pcr ◽

Expression Studies ◽

Gene Expression Studies ◽

Suitable Reference ◽

Selection Of

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Evaluation of Clostridium ljungdahlii DSM 13528 reference genes in gene expression studies by qRT-PCR

Journal of Bioscience and Bioengineering ◽

10.1016/j.jbiosc.2013.04.011 ◽

2013 ◽

Vol 116 (4) ◽

pp. 460-464 ◽

Cited By ~ 24

Author(s):

Juanjuan Liu ◽

Yang Tan ◽

Xiaohong Yang ◽

Xiaohua Chen ◽

Fuli Li

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Qrt Pcr ◽

Clostridium Ljungdahlii ◽

Expression Studies ◽

Gene Expression Studies

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Selection of suitable reference genes for gene expression studies in Staphylococcus capitis during growth under erythromycin stress

Molecular Genetics and Genomics ◽

10.1007/s00438-016-1197-9 ◽

2016 ◽

Vol 291 (4) ◽

pp. 1795-1811 ◽

Cited By ~ 1

Author(s):

Bintao Cui ◽

Peter M. Smooker ◽

Duncan A. Rouch ◽

Margaret A. Deighton

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Staphylococcus Capitis ◽

Expression Studies ◽

Gene Expression Studies ◽

Suitable Reference ◽

Selection Of

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Identification of suitable reference genes for gene expression studies of human serous ovarian cancer by real-time polymerase chain reaction

Analytical Biochemistry ◽

10.1016/j.ab.2009.07.022 ◽

2009 ◽

Vol 394 (1) ◽

pp. 110-116 ◽

Cited By ~ 67

Author(s):

Yan-Li Li ◽

Feng Ye ◽

Ying Hu ◽

Wei-Guo Lu ◽

Xing Xie

Keyword(s):

Gene Expression ◽

Ovarian Cancer ◽

Polymerase Chain Reaction ◽

Reference Genes ◽

Serous Ovarian Cancer ◽

Chain Reaction ◽

Expression Studies ◽

Polymerase Chain ◽

Gene Expression Studies ◽

Suitable Reference

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In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR

BMC Molecular Biology ◽

10.1186/1471-2199-8-47 ◽

2007 ◽

Vol 8 (1) ◽

pp. 47 ◽

Cited By ~ 105

Author(s):

Monika Jung ◽

Azizbek Ramankulov ◽

Jan Roigas ◽

Manfred Johannsen ◽

Martin Ringsdorf ◽

...

Keyword(s):

Gene Expression ◽

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Real Time Pcr ◽

Renal Cell ◽

Reference Genes ◽

Human Renal Cell Carcinoma ◽

Expression Studies ◽

Gene Expression Studies ◽

Suitable Reference

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Selection of suitable reference genes for abiotic stress-responsive gene expression studies in peanut by real-time quantitative PCR

Electronic Journal of Biotechnology ◽

10.1016/j.ejbt.2017.05.004 ◽

2017 ◽

Vol 28 ◽

pp. 76-86 ◽

Cited By ~ 6

Author(s):

Meijing He ◽

Shunli Cui ◽

Xinlei Yang ◽

Guojun Mu ◽

Huanying Chen ◽

...

Keyword(s):

Gene Expression ◽

Abiotic Stress ◽

Real Time ◽

Quantitative Pcr ◽

Reference Genes ◽

Real Time Quantitative Pcr ◽

Expression Studies ◽

Gene Expression Studies ◽

Suitable Reference ◽

Selection Of

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Gene expression studies in different genotypes of an ectomycorrhizal fungus require a high number of reliable reference genes

10.7287/peerj.preprints.2125v1 ◽

2016 ◽

Author(s):

Joske Ruytinx ◽

Tony Remans ◽

Jan V Colpaert

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Intraspecific Variability ◽

Standard Technique ◽

Housekeeping Genes ◽

Ectomycorrhizal Fungus ◽

Suillus Luteus ◽

Qrt Pcr ◽

Expression Studies ◽

Gene Expression Studies

Quantitative reverse transcription PCR (qRT-PCR) has become the standard technique for the expression analysis of a set of chosen genes of interest. The accuracy and reliability of qRT-PCR measurements strongly depends on the normalization with appropriate endogenous reference genes. In this study a set of candidate reference genes for the use in gene expression studies of a basidiomycete fungus, Suillus luteus, exposed to toxic concentrations of zinc or cadmium was identified, evaluated and validated. Seven candidate genes were selected from cDNA-AFLP as stably expressed and the algorithms geNorm and Normfinder were used to evaluate these genes alongside the traditionally used housekeeping genes (actin, tubulin) in different S. luteus isolates. The use of several S. luteus isolates revealed that each isolate has its own most stably expressed set of reference genes, regardless of the metal treatments, in casu metal exposures. Metal treatments had only a minor impact on the expression of the candidate reference genes. The validated reference genes outperform the in fungal research commonly used single, arbitrary chosen (“housekeeping”) genes in terms of reliability, and have the potential to be suitable reference genes when studying the effect of other environmental factors. A relatively high number of reference genes is required to correct for intraspecific variability when studying natural populations.

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