About: Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis     Goto   Sponge   NotDistinct   Permalink

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Description
  • 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 for the 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 occurrence of 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 barley caryopsis. This panel of genes could be used for other gene expression studies, as well as to optimize beta-amy1 analysis for study of the impact of beta-amy1 expression upon barley end-use quality.
  • 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 for the 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 occurrence of 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 barley caryopsis. This panel of genes could be used for other gene expression studies, as well as to optimize beta-amy1 analysis for study of the impact of beta-amy1 expression upon barley end-use quality. (en)
Title
  • Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis
  • Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis (en)
skos:prefLabel
  • Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis
  • Validation of the beta-amy1 Transcription Profiling Assay and Selection of Reference Genes Suited for a RT-qPCR Assay in Developing Barley Caryopsis (en)
skos:notation
  • RIV/00027006:_____/12:00002211!RIV13-MZE-00027006
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(GA521/07/1028), Z(MZE0002700604)
http://linked.open...iv/cisloPeriodika
  • 7
http://linked.open...vai/riv/dodaniDat
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  • 176900
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  • RIV/00027006:_____/12:00002211
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • REAL-TIME PCR; INTERNAL CONTROL GENES; MESSENGER-RNA; HOUSEKEEPING GENES; BETA-AMYLASE; EXPRESSION ANALYSIS; QUANTITATIVE PCR; QUALITY-CONTROL; ABIOTIC STRESS; IDENTIFICATION (en)
http://linked.open.../riv/klicoveSlovo
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  • US - Spojené státy americké
http://linked.open...ontrolniKodProRIV
  • [B89339056762]
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  • PLoS One
http://linked.open...in/vavai/riv/obor
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http://linked.open...vavai/riv/projekt
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  • 7
http://linked.open...iv/tvurceVysledku
  • Kučera, Ladislav
  • Ovesná, Jaroslava
  • Svobodová, I.
  • Milella, L.
  • Vaculová, K.
  • Štrymplová, Kamila
http://linked.open...ain/vavai/riv/wos
  • 000307045600044
http://linked.open...n/vavai/riv/zamer
issn
  • 1932-6203
number of pages
http://bibframe.org/vocab/doi
  • 10.1371/journal.pone.0041886
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