1. Introduction
2. Materials and Methods
2.1. Ethical Considerations
2.2. Blood Collection and DNA Extraction
2.3. Animal Genomic DNA Preparations
2.4. Preparation of Cloned Horse Transgenes as Reference Material
2.5. Quantitation of Transgenes Cloned into Plasmid Vector
2.6. Designing Primers and Probes for MFQPCR Detection
2.7. Preparation and Quantification of Control Samples
2.8. MFQPCR Detection Using Dynamic Array Integrated Fluid Circuits (IFCs)
2.9. Administration of EPO Transgene
3. Results and Discussion
3.1. Targeted Genes for Gene-Doping Control
3.2. Quantitation of Reference Materials (RMs)
3.3. Limit of Detection (LOD) of MFQPCR
3.4. MFQPCR-Amplification Specificity
3.5. Detection of the EPO Transgene After It Was Administered to a Horse
3.6. Casework Example
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene Name | Symbol | Function or Related Traits |
---|---|---|
creatine kinase, muscle | CKM | energy transduction |
erythropoietin | EPO | hematopoiesis |
fibroblast growth factor 2 | FGF2 | cell growth, morphogenesis, tissue repair |
follistatin | FST | muscle growth (antagonist of MSTN) |
growth hormone 1 | GH1 | growth control |
insulin like growth factor 1 | IGF1 | systemic body growth stimulation |
myostatin | MSTN | muscle growth (negative regulator) |
phosphoenolpyruvate carboxykinase 1 | PCK1 | regulation of gluconeogenesis |
pyruvate dehydrogenase kinase 4 | PDK4 | regulation of glucose and fatty acid metabolism |
peroxisome proliferator activated receptor delta | PPARD | oxidative (fat-burning) capacity |
vascular endothelial growth factor | VEGF | vasculogenesis, angiogenesis |
zinc finger and AT-hook domain containing | ZFAT | effect on body weight in thoroughbreds |
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