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Identification of Suitable Reference Genes for Gene Expression Normalization in Jatropha curcas L During Development and Under Stress Conditions Using Real Time Quantitative PCR BABT
Rocha,Antonio José; Maranhão,Paulo Abraão; Silva,Rafaela Oliveira; Pohl,Simone; Fonteles,Cristiane S.R.
Jatropha curcas L represent a potential source of raw material for the production of biodiesel. The aim this study was to find potential candidate reference genes in J. curcas tissues. Three softwares were utilized to verify which would be the most stable reference genes in qPCR assay: GeNorm, NormFinder and BestKeeper. The most stable reference genes in developing J. curcas seeds suggested by GeNorm were GAPDH, UCP, actin. However, the best combinations of stable genes in each tissue were identified separately under stress conditions: EF1-α, PP2A2 and GAPDH in total stress, however, in SA stress, four genes were required for normalization: PP2A2, EF1-α, GAPDH and PUB. In PEG stress, four genes also were required: PP2A2, EF1-α, GAPDH and PUB, while in NaCl...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Gene expression; RT-qPCR; Reference genes; Jatropha curcas L.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100404
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Evaluation of Reference Genes for Quantitative PCR in Four Tissues from Rabbits with Hypercholesterolaemia BABT
Zhang,Zhen; Wen,Bin; Xu,Yuan; Jiang,En-ze; Liu,Jia-yu; Zhu,Ke-li; Ning,Fang-yong; Du,Zhi-Heng; Bai,Xiu-Juan.
Abstract Rabbit with hypercholesterolaemia is an important model for studying cholesterol metabolism disease. This study aimed to evaluate the expression stability of nine reference genes for quantitative PCR (qPCR) analysis in adrenal gland, liver, spleen, and kidney tissue from rabbits with hypercholesterolaemia. In total, 30 male Harbin Large White (HLW) rabbits were fed a normal feed (n = 15) or a high cholesterol feed (n = 15) for 8 weeks to induce hypercholesterolaemia. Nine reference genes were verified by qPCR using cDNA extracted from rabbit tissue samples. For qPCR analysis, reference genes were evaluated using the RefFinder and GeNorm algorithms. Overall, seven rabbits with hypercholesterolaemia were identified based on body weight and total...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Evaluation; Reference genes; QPCR; Rabbits; Hypercholesterolaemia.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100428
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Real time PCR and importance of housekeepings genes for normalization and quantification of mRNA expression in different tissues BABT
Rebouças,Emanuela de Lima; Costa,José Jackson do Nascimento; Passos,Maria Juliane; Passos,José Renato de Sousa; Hurk,Robert van den; Silva,José Roberto Viana.
The aim of this review was to evaluate the importance of the real-time PCR (qRT-PCR) as a technique for mRNA expression analysis in different tissues. Real-time PCR is widely used for quantification of mRNA levels and is a fundamental tool for basic research, molecular medicine and biotechnology.Genes of references are expressed in a wide variety of tissues and cells with minimal variations in their expression levels, and thus are used to normalize data of mRNA quantification. Software programs, such as geNorm, BestKeeper and NormFinder, have been developed to perform the normalization of data, which help to choose the most stable reference gene. Several genes, such as GAPDH, β-actin, β-tubulin, PGK, UBQ, RPL-19 and 18S rRNA have been suggested as...
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR; Normalization; Reference genes; Gene expression; MRNA.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132013000100019
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Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis BJM
Chen,Chun; Xie,Tingna; Ye,Sudan; Jensen,Annette Bruun; Eilenberg,J ørgen.
Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pandora neoaphidis; Reference genes; Quantitative PCR.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259
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