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根据D03等11个微卫星引物单位点PCR扩增时优化的扩增条件(退火温度、MgCl2等)和11个微卫星引物所带有的荧光颜色、等位基因大小变异范围及反应的灵敏度,可将11个微卫星引物分成A组(D03、G04、B04和H08)、B组(A09、H11、A08和G01)和C组(CmrHr1.24、CmrHr2.30和CmrHr2.23)进行多元PCR扩增,结果显示各组内位点之间的分离清晰,这表明多元PCR可以大大提高微卫星检测仪器的使用效率和提高实验效果,在微卫星研究中是一种快速便捷的方法。
According to the optimized amplification conditions (annealing temperature, MgCl2, etc.) of 11 microsatellite primers at D03 and the fluorescence color, allelic size variation range and reaction sensitivity of 11 microsatellite primers, 11 microsatellite primers can be divided into A group (D03, G04, B04 and H08), B group (A09, H11, A08 and G01) and C group (CmrHr1.24, CmrHr2.30 and CmrHr2.23) The results showed that the separation between the sites within each group was clear, indicating that multiplex PCR can greatly improve the efficiency of microsatellite detection equipment and improve the experimental results, which is a quick and easy method in microsatellite research.