IUCN / SSC Cat Specialist Group - Digital Cat Library |
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| Jasa-Sangulin, L. | |
| Optimization of polymerase chain reaction for amplification of microsatellite markers of wildcat _Felis silvestris_ Schreber, 1777 | |
| 2017 Full Book | |
Genomic purity of European wildcat population is compromised by frequent hybridisation with domestic cats. Since the state Croatian wildcat population is unknown, it is necessary to carry out genotyping, i.e. amplify the microsatellite markers by polymerase chain reaction. In this work, optimization of PCR for amplification of 12 selected wildcat microsatellite markers was performed. An optimal amount of 10 ng DNA template and 0.2 æM of oligonucleotide primers was determined. Thermal cycling conditions were also optimized; optimal annealing temperature was 57øC and annealing and elongation time of 90 seconds. Finally, reaction conditions were further adjusted to perform multiplex PCR in order to save time and resources. Results of this work will be the basis of future amplification of microsatellite markers of wild and domestic cats in order to determine genomic purity of wildcat population in Croatia. |
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