Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci
Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci
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DOI：
10.1007/s00216-002-1683-2
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发表时间：
2003-01
影响因子：
4.3
通讯作者：
R. Schoske;P. Vallone;Christian M. Ruitberg;J. Butler
中科院分区：
文献类型：
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作者：
R. Schoske;P. Vallone;Christian M. Ruitberg;J. Butler
同时扩增DNA模板的多个区域通常使用聚合酶链反应（PCR）在称为多重PCR的过程中进行。一个有用的策略涉及设计，测试和优化多重PCR引物混合物将提出。其他多路设计协议侧重于引物的测试和优化，或嵌合引物的使用。通过对预测DNA低聚物熔融温度（Tm）和引物-二聚体相互作用的仔细检查，引物的设计可以减少获得一组平衡良好的扩增子所需的测试和优化量。本文构建的多重PCR引物混合物的测试和优化主要围绕着改变引物浓度，而不是测试多个引物组合。如果引物设计得当，只需要调整引物浓度，就可以得到一组平衡良好的扩增子。作为一个模型系统来说明这种多重设计方案，10个位点的多重（10plex） Y染色体短串联重复（STR）测定被使用。
The simultaneous amplification of multiple regions of a DNA template is routinely performed using the polymerase chain reaction (PCR) in a process termed multiplex PCR. A useful strategy involving the design, testing, and optimization of multiplex PCR primer mixtures will be presented. Other multiplex design protocols have focused on the testing and optimization of primers, or the use of chimeric primers. The design of primers, through the close examination of predicted DNA oligomer melting temperatures (Tm) and primer–dimer interactions, can reduce the amount of testing and optimization required to obtain a well-balanced set of amplicons. The testing and optimization of the multiplex PCR primer mixture constructed here revolves around varying the primer concentrations rather than testing multiple primer combinations. By solely adjusting primer concentrations, a well-balanced set of amplicons should result if the primers were designed properly. As a model system to illustrate this multiplex design protocol, a 10-loci multiplex (10plex) Y chromosome short tandem repeat (STR) assay is used.