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《Chinese Journal of Biochemistry and Molecular Biology》 2009-12
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Identification of Suitable Reference for Quantitative RT-PCR Assays of miR-103 in Pig Tissues

LI Guo-Xi1),2),NING Xiao-Min1),LI Xin-Jian1),2),WU Zong-Song1),YANG Gong-She1)(1)Laboratory of Animal Fat Deposition and Muscle Development,Northwest A&F University,Yangling 712100,Shaanxi,China;2)College of Animal Husbandry & Veterinary Science,Henan Agricultural University,Zhengzhou 450002,Henan,China)  
Quantitative real-time PCR(qRT-PCR) is one of the common methods for determining the miRNA transcriptions.The proper normalization of the qRT-PCR results using appropriate references is critical for the accuracy of this method.In order to identify an optimal reference for miR-103 qRT-PCR assays in normal pig tissues,three candidate references,miR-196,U6 snRNA and total RNA,were assessed by qRT-PCR with the geNorm algorithm and the amplification efficiency of experimental validation.The accuracies of the three candidate references were compared based on the absolute quantification of miR-103 with the control.The results showed that miR-196 outperformed than U6 snRNA and total RNA both in the stability and the amplification efficiency.The relative quantization normalized to miR-196 was highly correlated to the miR-103 absolute amount.We found that miR-103 was most abundantly in the cerebrum,moderately transcribed in the stomach,cerebellum,small intestine,heart,liver and pancreas,and weak in the lung,spleen and crura muscles.These results indicated that miR-196 was a suitable reference for assaying relative levels of miR-103 by qRT-PCR in normal pig tissues.
【Fund】: 国家高技术研究发展计划(863计划)(No.2006AA10Z138)~~
【CateGory Index】: Q953
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