Selection of Reference Genes for Normalization of Gene Expression Data in Blood of Machado-Joseph Disease/Spinocerebellar Ataxia Type 3 (MJD/SCA3) Subjects

Ferreira, Ana F.; Raposo, Mafalda; Vasconcelos, João; Costa, Maria do Carmo; Lima, Manuela

doi:10.1007/s12031-019-01374-0

Cited by 7 publications

(10 citation statements)

References 19 publications

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“…For example, in different heart and disease conditions, PPIA is recognized by ReFinder as the best reference gene in different skeletal muscles of mice, and it ranked first for human endometrial cancer [52,53]. PPIB is believed as the optimal reference gene in analyzing the blood of Machado-Joseph disease (MJD) patients [54].…”

Section: Discussionmentioning

confidence: 99%

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

et al. 2020

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Background Before studying gene expression of different organisms, it is important to determine the best reference gene. At present, the most accurate method of detecting gene expression is quantitative real-time PCR (RT-qPCR). With this method, reference genes that are stable in different biological systems and under different conditions can be obtained. Toona ciliata Roem (T. ciliata). is a valuable and fast-growing timber specie. In this study, 20 reference genes were identified using RT-qPCR, as a primary prerequisite for future gene expression analysis. Four different methods, geNorm, NormFinder, BestKeeper, and RankAggreg were used to evaluate the expression stability of the 20 candidate reference genes in various tissues under different conditions. Results The experimental results showed that TUB-α was the most stably expressed reference gene across all samples and UBC17 was the most stable in leaves and young stems under Hypsipyla robusta (H. robusta) and methyl jasmonate (MeJA) treatments. In addition, PP2C59 and UBC5B were the best-performing genes in leaves under H. robusta treatment, while HIS1 and ACT7 were the best reference genes in young stems. The two best reference genes were 60S-18 and TUB-α after treatment at 4 °C. The expression of HIS6 and MUB1 was the most stable under PEG6000 treatment. The accuracy of the selected reference genes was verified using the transcription factor MYB3 (TcMYB3) gene. Conclusions This is the first report to verify the best reference genes for normalizing gene expression in T. ciliata under different conditions, which will facilitate future elucidation of gene regulations in this species.

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Section: Discussionmentioning

confidence: 99%

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

et al. 2020

View full text Add to dashboard Cite

show abstract

“…For example, in different heart and disease conditions, PPIA is recognized by ReFinder as the best reference gene in different skeletal muscles of mice, and it ranked rst for human endometrial cancer [52,53]. PPIB is believed as the optimal reference gene in analyzing the blood of Machado-Joseph disease (MJD) patients [54].…”

Section: Discussionmentioning

confidence: 99%

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

Song

Mao

Duan

et al. 2020

Preprint

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Background: Before studying gene expression of different organisms, it is important to determine the best reference gene. At present, the most accurate method of detecting gene expression is quantitative real-time PCR (RT-qPCR). With this method, reference genes that are stable in different biological systems and under different conditions can be obtained. Toona ciliata Roem (T. ciliata). is a valuable and fast-growing timber specie. In this study, 20 reference genes were identified using RT-qPCR, as a primary prerequisite for future gene expression analysis. Four different methods, geNorm, NormFinder, BestKeeper, and RankAggreg were used to evaluate the expression stability of the 20 candidate reference genes in various tissues under different conditions. Results: The experimental results showed that TUB-α was the most stably expressed reference gene across all samples and UBC17 was the most stable in leaves and young stems under Hypsipyla robusta (H. robusta) and methyl jasmonate (MeJA) treatments. In addition, PP2C59 and UBC5B were the best-performing genes in leaves under H. robusta treatment, while HIS1 and ACT7 were the best reference genes in young stems. The two best reference genes were 60S-18 and TUB-α after treatment at 4 °C. The expression of HIS6 and MUB1 was the most stable under PEG6000 treatment. The accuracy of the selected reference genes was verified using the transcription factor MYB3 (TcMYB3) gene. Conclusions: This is the first report to verify the best reference genes for normalizing gene expression in T. ciliata under different conditions, which will facilitate future elucidation of gene regulations in this species.

show abstract

“…For example, in different heart and disease conditions, PPIA is recognized by ReFinder as the best reference gene in different skeletal muscles of mice, and it is ranked rst for human endometrial cancer [50,51]. PPIB is believed as more optimal reference gene in analyzing the blood of Machado-Joseph disease (MJD) patients [52].…”

Section: Discussionmentioning

confidence: 99%

Selection and Validation of Reference Genes for Measuring Gene Expression in Toona Ciliata under Different Experimental Conditions by Quantitative Real-Time PCR Analysis

Song

Mao

Duan

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Before studying the gene expression of different organisms, it is important to determine the best reference gene. At present, the most accurate method for detecting gene expression is quantitative real-time PCR(RT-qPCR). By using this method, reference genes that are stable in different biological systems and under different conditions can be obtained. Toona ciliata Roem (T. ciliata). is a valuable and fast-growing timber species. In this study, 20 reference genes were identified through RT-qPCR, as a primary prerequisite for future gene expression analysis. Four different methods, geNorm, NormFinder, BestKeeper, and RankAggreg were used to evaluate the stability of expression of 20 candidate reference genes in various tissues under different conditions. Results: The experimental results showed that TUB-α was the most stably expressed reference gene across all samples; UBC17 was found to be the most stable in leaves & young stems under Hypsipyla robusta (H. robusta) and methyl jasmonate (MeJA) treatment. In addition, under H. robusta treatment, PP2C59 and UBC5B were the best-performing genes in leaves, while HIS1 and ACT7 were the best reference genes in young stems. Under low temperature (4℃) treatment, the two best reference genes were 60S-18 and TUB-α. The expression of HIS6 and MUB1 was the most stable under PEG6000 treatment. The accuracy of the selected reference genes was verified using transcription factor MYB3(TcMYB3) genes. Conclusions: This is the first report to verify the best reference genes for normalizing gene expression in T. ciliata under different conditions, which will facilitate the future elucidation of gene regulations in this species.

show abstract

Selection of Reference Genes for Normalization of Gene Expression Data in Blood of Machado-Joseph Disease/Spinocerebellar Ataxia Type 3 (MJD/SCA3) Subjects

Cited by 7 publications

References 19 publications

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

Selection and validation of reference genes for measuring gene expression in Toona ciliata under different experimental conditions by quantitative real-time PCR analysis

Selection and Validation of Reference Genes for Measuring Gene Expression in Toona Ciliata under Different Experimental Conditions by Quantitative Real-Time PCR Analysis

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