两种扩增仪对联合毛细管电泳技术的聚合酶链式反应的影响

Effect of the polymerase chain reaction combined capillary electrophoresis on two amplification instruments

目的使用两种扩增仪进行常规扩增和毛细管电泳,观察两种扩增仪对结果的影响以及有何不同。方法运用Power Plex试剂盒对DNA在Applied Biosystems 9700和Hema 9700两种扩增仪上进行扩增,扩增包括对血卡进行直接扩增和对提取后的DNA进行扩增,然后在Applied Biosystems 3130XL上进行毛细管电泳检测;使用同一DNA进行5次重复试验进行扩增仪的稳定性检测;使用7种非人样本DNA进行种属特异性检测;运用不同稀释度的2800阳性标准品进行灵敏度检测;对使用两种扩增仪扩增检测后的所有结果进行分析、讨论和评价。结果使用两种扩增仪进行扩增检测后,其分型结果在两种扩增仪之间没有明显差别,其分型结果均稳定准确且在两种扩增仪上呈现一致性,种属特异性检测结果一致,灵敏度检测结果一致。结论两种扩增仪在进行常规扩增检测时,其结果在两种扩增仪之间没有明显差别,其结果呈现出一致性,两种扩增仪均可应用于常规扩增。

Objective To observe the effects and differences between the two amplification instruments combined capillary electrophoresis. Methods DNA extracted were amplified with the optimized PCR method and cycling protocol using Powerplex 21 kit on the Applied Biosystems 9700 and HEMA 9700 instrument, including direct amplification and DNA amplification, and then genotyped using Applied Biosystems 3130xl. Same DNA test was repeated five times to test the stability of the instrument, seven kinds of non-human DNA samples were detected and 9947 was used as positive standard to test the sensitivity. Then genotyping results were validated and discussed. Results Genotyping results between the two instruments had no significant differences, genotyping results were stable and accurate and showed consistency in species-specific detection and sensitivity detection between the two amplification instruments. Conclusions There is no significant difference between the two amplification instruments of the routine amplification test and the results were consistent. The results showed that the two amplification instruments could be used in routine amplification.