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目的优化和建立一套快速聚合酶链式反应(PCR)扩增体系和程序,缩短扩增时间,提高短串联重复序列分型的速率和办案效率。方法运用AmpFLSTR?Identifiler?Plus试剂盒和7种快速扩增酶进行筛选和优化,最后对筛选出的酶优化其扩增体系和扩增程序,并对几种常规检材中提取的DNA进行扩增和检测,然后对其分型结果进行验证和讨论。结果最终优化的扩增程序由常规的3 h左右缩短至24 min,DNA检验结果与常规方法一致。结论应用优化后的快速PCR扩增体系可以成功准确扩增DNA样本,显著提高DNA分型速率。
Objective To optimize and establish a rapid polymerase chain reaction (PCR) amplification system and program to shorten the amplification time and improve the rate of short tandem repeat typing and case handling efficiency. Methods AmpFLSTR? Identifiler? Plus kit and seven kinds of rapid amplification enzyme screening and optimization, and finally the selected enzyme to optimize its amplification system and amplification program, and several conventional samples extracted DNA amplification Increase and test, and then verify the results of their typing and discussion. Results The final optimized amplification procedure was shortened from about 3 h to 24 min, and the DNA test results were consistent with the conventional methods. Conclusion The optimized rapid PCR amplification system can successfully and accurately amplify DNA samples and significantly improve the DNA typing rate.