Počet záznamů: 1
Evaluation of reference genes for reverse transcription quantitative real-time PCR (RT-qPCR) studies in Silene vulgaris considering the method of cDNA preparation
- 1.
SYSNO ASEP 0477711 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Evaluation of reference genes for reverse transcription quantitative real-time PCR (RT-qPCR) studies in Silene vulgaris considering the method of cDNA preparation Tvůrce(i) Koloušková, Pavla (UEB-Q)
Stone, James D. (UEB-Q)
Štorchová, Helena (UEB-Q) RID, ORCIDCelkový počet autorů 3 Číslo článku e0183470 Zdroj.dok. PLoS ONE. - : Public Library of Science - ISSN 1932-6203
Roč. 12, č. 8 (2017)Poč.str. 14 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova CYTOPLASMIC MALE-STERILITY ; SUITABLE REFERENCE GENES ; INTERNAL CONTROL GENES Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Plant sciences, botany CEP LH15075 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy GA16-09220S GA ČR - Grantová agentura ČR Institucionální podpora UEB-Q - RVO:61389030 UT WOS 000407856600140 EID SCOPUS 85027705328 DOI 10.1371/journal.pone.0183470 Anotace 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. Pracoviště Ústav experimentální botaniky Kontakt David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Rok sběru 2018
Počet záznamů: 1