Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis

Kód výsledku v IS VaVaI

RIV/00027006:_____/12:00002211 - isvavai.cz

Nalezeny alternativní kódy

RIV/25328859:_____/12:#0000654

Výsledek na webu

http://dx.doi.org/10.1371/journal.pone.0041886

DOI - Digital Object Identifier

10.1371/journal.pone.0041886

Jazyk výsledku

angličtina

Název v původním jazyce

Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis

Popis výsledku v původním jazyce

Reverse transcription coupled with real-time quantitative PCR (RT-qPCR) is a frequently used method for gene expression profiling. Reference genes (RGs) are commonly employed to normalize gene expression data. A limited information exist on the gene expression and profiling in developing barley caryopsis. Expression stability was assessed by measuring the cycle threshold (Ct) range and applying both the GeNorm (pair-wise comparison of geometric means) and Normfinder (model-based approach) principles forthe calculation. Here, we have identified a set of four RGs suitable for studying gene expression in the developing barley caryopsis. These encode the proteins GAPDH, HSP90, HSP70 and ubiquitin. We found a correlation between the frequency of occurrenceof a transcript in silico and its suitability as an RG. This set of RGs was tested by comparing the normalized level of beta-amylase (beta-amy1) transcript with directly measured quantities of the BMY1 gene product in the developing barl

Název v anglickém jazyce

Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis

Popis výsledku anglicky

Reverse transcription coupled with real-time quantitative PCR (RT-qPCR) is a frequently used method for gene expression profiling. Reference genes (RGs) are commonly employed to normalize gene expression data. A limited information exist on the gene expression and profiling in developing barley caryopsis. Expression stability was assessed by measuring the cycle threshold (Ct) range and applying both the GeNorm (pair-wise comparison of geometric means) and Normfinder (model-based approach) principles forthe calculation. Here, we have identified a set of four RGs suitable for studying gene expression in the developing barley caryopsis. These encode the proteins GAPDH, HSP90, HSP70 and ubiquitin. We found a correlation between the frequency of occurrenceof a transcript in silico and its suitability as an RG. This set of RGs was tested by comparing the normalized level of beta-amylase (beta-amy1) transcript with directly measured quantities of the BMY1 gene product in the developing barl

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

GE - Šlechtění rostlin

OECD FORD obor

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Projekt

GA521/07/1028: Analýza exprese genu kódujícího enzym beta-amylázu v zrnu ječmene ve vztahu ke strukturálnímu polymorfismu genu a kvalitativním vlastnostem zrna

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2012

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

PLoS One

ISSN

1932-6203

e-ISSN

—

Svazek periodika

7

Číslo periodika v rámci svazku

7

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

1

Strana od-do

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Kód UT WoS článku

000307045600044

EID výsledku v databázi Scopus

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