Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.

The real time quantitative reverse transcription PCR (qRT-PCR) is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and strin...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Ruby Chandna, Rehna Augustine, Naveen C Bisht
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2a57c01520bd495c90a3237b52e28570
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2a57c01520bd495c90a3237b52e28570
record_format	dspace
spelling	oai:doaj.org-article:2a57c01520bd495c90a3237b52e285702021-11-18T07:19:06ZEvaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.1932-620310.1371/journal.pone.0036918https://doaj.org/article/2a57c01520bd495c90a3237b52e285702012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22606308/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The real time quantitative reverse transcription PCR (qRT-PCR) is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes), ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes), in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.Ruby ChandnaRehna AugustineNaveen C BishtPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 5, p e36918 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ruby Chandna Rehna Augustine Naveen C Bisht Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
description	The real time quantitative reverse transcription PCR (qRT-PCR) is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes), ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes), in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.
format	article
author	Ruby Chandna Rehna Augustine Naveen C Bisht
author_facet	Ruby Chandna Rehna Augustine Naveen C Bisht
author_sort	Ruby Chandna
title	Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
title_short	Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
title_full	Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
title_fullStr	Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
title_full_unstemmed	Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
title_sort	evaluation of candidate reference genes for gene expression normalization in brassica juncea using real time quantitative rt-pcr.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/2a57c01520bd495c90a3237b52e28570
work_keys_str_mv	AT rubychandna evaluationofcandidatereferencegenesforgeneexpressionnormalizationinbrassicajunceausingrealtimequantitativertpcr AT rehnaaugustine evaluationofcandidatereferencegenesforgeneexpressionnormalizationinbrassicajunceausingrealtimequantitativertpcr AT naveencbisht evaluationofcandidatereferencegenesforgeneexpressionnormalizationinbrassicajunceausingrealtimequantitativertpcr
_version_	1718423586256977920

Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.

Ejemplares similares