Validation of reference genes for gene expression analysis in Valsa mali var. mali using real-time quantitative PCR

2013 ◽  
Vol 29 (9) ◽  
pp. 1563-1571 ◽  
Author(s):  
Zhiyuan Yin ◽  
Xiwang Ke ◽  
Dingxuan Huang ◽  
Xiaoning Gao ◽  
Ralf T. Voegele ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Expression Analysis ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Real Time Quantitative Pcr ◽  
Valsa Mali

scholarly journals Identification and validation of quantitative real-time reverse transcription PCR reference genes for gene expression analysis in teak (Tectona grandis L.f.)

2014 ◽  
Vol 7 (1) ◽  
pp. 464 ◽  
Author(s):  
Esteban Galeano ◽  
Tarcísio Vasconcelos ◽  
Daniel Ramiro ◽  
Valentina de De Martin ◽  
Helaine Carrer
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Tectona Grandis ◽  
Reverse Transcription Pcr ◽  
Tectona Grandis L.F

Validation of reference genes for accurate normalization of gene expression for real time-quantitative PCR in strawberry fruits using different cultivars and osmotic stresses

Gene ◽  
2015 ◽  
Vol 554 (2) ◽  
pp. 205-214 ◽  
Author(s):  
Vanessa Galli ◽  
Joyce Moura Borowski ◽  
Ellen Cristina Perin ◽  
Rafael da Silva Messias ◽  
Julia Labonde ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Reference Genes ◽  
Real Time Quantitative Pcr ◽  
Accurate Normalization

scholarly journals Establishing reference genes for use in real-time quantitative PCR analysis of early equine embryos

2011 ◽  
Vol 23 (2) ◽  
pp. 353 ◽  
Author(s):  
Damien B. B. P. Paris ◽  
Ewart W. Kuijk ◽  
Bernard A. J. Roelen ◽  
Tom A. E. Stout
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Embryo Development ◽  
Reference Genes ◽  
Stable Expression ◽  
Pcr Analysis ◽  
Blastocyst Development ◽  
Real Time Quantitative Pcr

Real-time quantitative PCR (qPCR) is invaluable for investigating changes in gene expression during early development, since it can be performed on the limited quantities of mRNA contained in individual embryos. However, the reliability of this method depends on the use of validated stably expressed reference genes for accurate data normalisation. The aim of the present study was to identify and validate a set of reference genes suitable for studying gene expression during equine embryo development. The stable expression of four carefully selected reference genes and one developmentally regulated gene was examined by qPCR in equine in vivo embryos from morula to expanded blastocyst stage. SRP14, RPL4 and PGK1 were identified by geNorm analysis as stably expressed reference genes suitable for data normalisation. RPL13A expression was less stable and changed significantly during the period of development examined, rendering it unsuitable as a reference gene. As anticipated, CDX2 expression increased significantly during embryo development, supporting its possible role in trophectoderm specification in the horse. In summary, it was demonstrated that evidence-based selection of potential reference genes can reduce the number needed to validate stable expression in an experimental system; this is particularly useful when dealing with tissues that yield small amounts of mRNA. SRP14, RPL4 and PGK1 are stable reference genes suitable for normalising expression for genes of interest during in vivo morula to expanded blastocyst development of horse embryos.

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

2017 ◽  
Vol 28 ◽  
pp. 76-86 ◽  
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

scholarly journals Evaluation of putative reference genes for quantitative real-time PCR normalization inLilium regaleduring development and under stress

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1837 ◽  
Author(s):  
Qiang Liu ◽  
Chi Wei ◽  
Ming-Fang Zhang ◽  
Gui-Xia Jia
Keyword(s):  
Gene Expression ◽  
Heat Stress ◽  
Real Time ◽  
Quantitative Pcr ◽  
Biotic Stress ◽  
Reference Genes ◽  
Developmental Stages ◽  
Real Time Quantitative Pcr ◽  
The Stability ◽  
Experimental Treatments

Normalization to reference genes is the most common method to avoid bias in real-time quantitative PCR (qPCR), which has been widely used for quantification of gene expression. Despite several studies on gene expression,Lilium, and particularlyL. regale, has not been fully investigated regarding the evaluation of reference genes suitable for normalization. In this study, nine putative reference genes, namely18S rRNA,ACT,BHLH,CLA,CYP,EF1,GAPDH,SANDandTIP41, were analyzed for accurate quantitative PCR normalization at different developmental stages and under different stress conditions, including biotic (Botrytis elliptica), drought, salinity, cold and heat stress. All these genes showed a wide variation in their Cq (quantification Cycle) values, and their stabilities were calculated by geNorm, NormFinder and BestKeeper. In a combination of the results from the three algorithms,BHLHwas superior to the other candidates when all the experimental treatments were analyzed together;CLAandEF1were also recommended by two of the three algorithms. As for specific conditions,EF1under various developmental stages,SANDunder biotic stress,CYP/GAPDHunder drought stress, andTIP41under salinity stress were generally considered suitable. All the algorithms agreed on the stability ofSANDandGAPDHunder cold stress, while onlyCYPwas selected under heat stress by all of them. Additionally, the selection of optimal reference genes under biotic stress was further verified by analyzing the expression level ofLrLOXin leaves inoculated withB. elliptica. Our study would be beneficial for future studies on gene expression and molecular breeding ofLilium.

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