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Evaluation of reference genes for reverse transcription quantitative real-time PCR (RT-qPCR) studies in Silene vulgaris considering the method of cDNA preparation

SYSNO ASEP	0477711
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Evaluation of reference genes for reverse transcription quantitative real-time PCR (RT-qPCR) studies in Silene vulgaris considering the method of cDNA preparation
Author(s)	Koloušková, Pavla (UEB-Q) Stone, James D. (UEB-Q) Štorchová, Helena (UEB-Q)_{RID, ORCID}
Number of authors	3
Article number	e0183470
Source Title	PLoS ONE. - : Public Library of Science - ISSN 1932-6203 Roč. 12, č. 8 (2017)
Number of pages	14 s.
Language	eng - English
Country	US - United States
Keywords	CYTOPLASMIC MALE-STERILITY ; SUITABLE REFERENCE GENES ; INTERNAL CONTROL GENES
Subject RIV	EB - Genetics ; Molecular Biology
OECD category	Plant sciences, botany
R&D Projects	LH15075 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	GA16-09220S GA ČR - Czech Science Foundation (CSF)
Institutional support	UEB-Q - RVO:61389030
UT WOS	000407856600140
EID SCOPUS	85027705328
DOI	10.1371/journal.pone.0183470
Annotation	Accurate gene expression measurements are essential in studies of both crop and wild plants. Reverse transcription quantitative real-time PCR (RT-qPCR) has become a preferred tool for gene expression estimation. A selection of suitable reference genes for the normalization of transcript levels is an essential prerequisite of accurate RT-qPCR results. We evaluated the expression stability of eight candidate reference genes across roots, leaves, flower buds and pollen of Silene vulgaris (bladder campion), a model plant for the study of gynodioecy. As random priming of cDNA is recommended for the study of organellar transcripts and poly(A) selection is indicated for nuclear transcripts, we estimated gene expression with both random-primed and oligo(dT)-primed cDNA. Accordingly, we determined reference genes that perform well with oligo(dT)- and random-primed cDNA, making it possible to estimate levels of nucleus-derived transcripts in the same cDNA samples as used for organellar transcripts, a key benefit in studies of cyto-nuclear interactions. Gene expression variance was estimated by RefFinder, which integrates four different analytical tools. The SvACT and SvGAPDH genes were the most stable candidates across various organs of S. vulgaris, regardless of whether pollen was included or not.
Workplace	Institute of Experimental Botany
Contact	David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469
Year of Publishing	2018

Number of the records: 1