Identification of normalization factors for quantitative real-time RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai

Qiu, Reng; Sun, Bo; Fang, Shasha; Sun, Li; Xiao, Lianchun

doi:10.1007/s00343-013-2221-0

Cited by 12 publications

(2 citation statements)

References 39 publications

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“…Notably, a recent study with different abalone species (red abalone H. rufescens) reported that ACTB was the least stable reference in the hepatopancreas (Lόpez-Landavery et al 2014). The unstable feature of the ACTB gene was also reported in this abalone species during bacterial challenge (Qiu et al 2013). The significantly different basal levels of the ACTB transcripts across different tissues in aquatic animals might also be, at least in part, supportive of our finding with this abalone species (Cho et al 2011;Kim et al 2008).…”

Section: Pairwise Variation (V)mentioning

confidence: 99%

Evaluation of reference genes for RT-qPCR study in abalone Haliotis discus hannai during heavy metal overload stress

Lee

Nam

2016

Fish Aquatic Sci

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Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ΔCT method). Results: Expression stability in the gill subset was determined asOn the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as β-tubulin (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts.Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

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Section: Pairwise Variation (V)mentioning

confidence: 99%

Evaluation of reference genes for RT-qPCR study in abalone Haliotis discus hannai during heavy metal overload stress

Lee

Nam

2016

Fish Aquatic Sci

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“…Comprehensive understanding genes and its expression involved in the development and ontogeny would be a fundamental requirement for all these breeding investigations with regard to evaluate developmental characteristics and early performances of newly developed breeds. Evaluation of reference genes for RT-qPCR normalization in Haliotis species has been reported with respect to type of tissues and experimental challenges using toxicants and bacteria (Wan et al, 2011;Qiu et al, 2013;López-Landavery, 2014;Lee and Nam, 2016a…”

mentioning

confidence: 99%

Assessment of Suitable Reference Genes for RT-qPCR Normalization with Developmental Samples in Pacific Abalone Haliotis discus hannai

Lee

Park

Nam

2019

J Anim Reprod Biotechnol

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Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation

Zhao

Chen

Wang

et al. 2014

Chin. J. Ocean. Limnol.

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Identification of normalization factors for quantitative real-time RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai

Cited by 12 publications

References 39 publications

Evaluation of reference genes for RT-qPCR study in abalone Haliotis discus hannai during heavy metal overload stress

Evaluation of reference genes for RT-qPCR study in abalone Haliotis discus hannai during heavy metal overload stress

Assessment of Suitable Reference Genes for RT-qPCR Normalization with Developmental Samples in Pacific Abalone Haliotis discus hannai

Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation

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