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Setting up a gene expression study for tissue cells by method of quantitative real-time PCR
Dušanić, Daliborka (Author), Bolha, Luka (Author), Narat, Mojca (Author), Oven, Irena (Author)

URLURL - Presentation file, Visit http://aas.bf.uni-lj.si/zootehnika/100-2012/PDF/100-2012-1-19-28.pdf This link opens in a new window

Abstract
Gene expression analysis by quantitative reverse transcription polymerase chain reaction (RT-qPCR) allows accurate and sensitive measurment of gene expression levels. However, a series of steps needs to be taken to ensure the relevance, accuracy, correct interpretation and repeatability of the RT-qPCR experiment. We describe here a simple experiment of determining relative gene expression for caspase 8 gene in chicken chondrocytes treated with an apoptosis inducing compound 5-fluorouracil. We use this example to point out some important guidelines in setting up a gene expression study in tissue cells, analyzing and interpreting the results and reporting on the findings.

Language:English
Keywords:molecular genetics, RT-qPCR, gene expression, tissue cells
Work type:Not categorized (r6)
Tipology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Year:2012
Number of pages:str. 19-28
Numbering:Letn. 100, št. 1
UDC:575
ISSN on article:1581-9175
COBISS.SI-ID:3081864 Link is opened in a new window
Views:572
Downloads:185
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Record is a part of a journal

Title:Acta agriculturae Slovenica
Shortened title:Acta agric. Slov.
Publisher:Biotehniška fakulteta
ISSN:1581-9175
COBISS.SI-ID:213840640 This link opens in a new window

Secondary language

Language:Slovenian
Abstract:
Analiza izražanja genov z metodo reverznega pomnoževanja in kvantitativne verižne reakcije s polimerazo v realnem času (RT-qPCR) omogoča natančno in občutljivo določanje nivoja izražanja tarčnih genov. Vendar je za zagotavljanje točnosti, natančne interpretacije in ponovljivosti eksperimenta, predvsem pa relevantnosti rezultatov, pomembno natančno poznavanje korakov postopka. V tem prispevku opisujemo enostaven poskus, v katerem smo v celični liniji kokošjih hondrocitov, ki smo jih tretirali s spojino 5-fluorouracil, ki sproži apoptozo, določili relativno izražanje gena, ki kodira encim kaspazo 8.Ta primer smo uporabili za izpostavitev nekaterih pomembnih smernic pri oblikovanju študije izražanja genov v tkivnih celicah, analizi in interpretaciji rezultatov ter oblikovanju zaključkov.

Keywords:molekularna genetika, kvantitativni PCR v realnem času, genetsko izražanje, tkivne celice

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