Measured and compared against the expression level of the target Particular transcript, the input copy number is calculated, whereasДЇor relative quantification, the expression of a reference gene is (RT-qPCR) is an extremely sensitive method for studying absolute or These results may affect the choice of reference genes in future studies that use RTвЂ‘qPCR analysis of target genes in experimental conditions, such as mice with KYDS infected with influenza A virus.ДЄt present, reverse transcription-quantitative PCR In contrast, the least stable genes in all 4В tissues were GAPDH and ОІ2m. From the 4В algorithms, taking into account the joint analyses of the ranking order outputs, the 2В genes Ywhae and Nedd8 were identified to be the most stable for mice with KYDS following infection with A/H1N1 virus. The results were authenticated through extended experimental settings using a group of 10В samples, parallel to 3 additional innate immune systemвЂ‘associated genes of the host, TLR3, TLR7 and RIGвЂ‘I, which were also analyzed using the same algorithms. For analysis, geNorm, BestKeeper, NormFinder, and BioвЂ‘Rad Maestro в„ў statistical programs were used for the estimation of the stability of the reference genes. KYDS mice were inoculated with A/H1N1 virus or a mock control.

In the present study, analysis of 10 reference genes (ActвЂ‘ОІ, ОІ2m, GAPDH, GusОІ, TubО±, Grcc10, Eif4h, Rnf187, Nedd8 and Ywhae) was performed across a set of 4 tissue types: Lung heart liver and kidney. To accurately estimate the relative expression of genes in cells from mice with KYDS in response to infection with influenza A virus subtype H1N1 (A/H1N1) virus using RTвЂ‘qPCR, it is necessary to identify suitable reference genes.

ReverseвЂ‘transcriptionвЂ‘quantitative PCR (RTвЂ‘qPCR) is considered an extremely sensitive technique used for absolute and relative quantification of target genes transcript levels. A stable reference gene is essential as an internal control for molecular biology research of this condition.

However, the specific molecular mechanisms underlying this disease remain unclear. KidneyвЂ‘yang deficiency syndrome (KYDS) infected with the influenza virus is a suitable model to imitate a population at highвЂ‘risk to influenza infection with a high rate of morbidity and mortality.

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