Identification of endogenous normalizing genes for expression studies in inguinal ring tissue for scrotal hernias in pigs

PloS One
William Raphael LorenzettiMônica Corrêa Ledur

Abstract

The use of reference genes is required for relative quantification in gene expression analysis and since the stability of these genes could be variable depending on the experimental design, it has become indispensable to test the reliability of endogenous genes. Therefore, this study evaluated 10 reference candidate genes in two different experimental conditions in order to obtain stable genes to be used as reference in expression studies related to scrotal hernias in pigs. Two independent experiments were performed: one with 30 days-old MS115 pigs and the other with 60 days-old Landrace pigs. The inguinal ring/canal was collected, frozen and further submitted to real-time PCR analysis (qPCR). For the reference genes stability evaluation, four tools were used: GeNorm in the SLqPCR, BestKeeper, NormFinder and Comparative CT. A general ranking was generated using the BruteAggreg function of R environment. In this study, the RPL19 was one of the most reliable endogenous genes for both experiments. The breed/age effects influenced the expression stability of candidate reference genes evaluated in the inguinal ring of pigs. Therefore, this study reinforces the importance of evaluating the stability of several endogenous genes previo...Continue Reading

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Citations

Oct 5, 2019·Molecular Biology Reports·Caroline Michele Marinho MarcianoMônica Corrêa Ledur

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Methods Mentioned

BETA
PCR
protein folding

Software Mentioned

Excel
Bestkeeper
SLqPCR
R environment
Primer
RefFinder
BruteAggreg
DeltaCt
- Blast
Comparative Ct

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