Multiplex allele-specific target amplification based on PCR suppression
Multiplex allele-specific target amplification based on PCR suppression
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DOI：
10.1073/pnas.98.1.206
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发表时间：
2001-01-02
影响因子：
11.1
通讯作者：
Cantor, CR
中科院分区：
文献类型：
--
作者：
Broude, NE;Zhang, LG;Cantor, CR
我们已经开发了一种基于PCR抑制的多重PCR策略。PCR抑制允许DNA靶扩增，每个靶只有一个序列特异性引物和所有靶共有的第二个引物。因此，n重PCR将仅需要n + 1个引物。我们已经证明了在14重PCR中靶序列的均匀、有效的扩增。抑制PCR的高特异性还提供了具有等位基因特异性的多重扩增。多重PCR用于开发用于对来自囊性纤维化受影响个体的DMA样品进行基因分型的测定。新方法大大简化了引物设计，显著提高了PCR多重化水平，并降低了总体引物成本。此外，该测定更易于自动化，因此适用于高通量遗传诊断。
We have developed a strategy for multiplex PCR based on PCR suppression. PCR suppression allows DNA target amplification with only one sequence-specific primer per target and a second primer that is common for all targets. Therefore, an n-plex PCR would require only n + 1 primers. We have demonstrated uniform, efficient amplification of targeted sequences in 14-plex PCR. The high specificity of suppression PCR also provides multiplexed amplification with allele specificity. Multiplexed PCR was used to develop assays for genotyping DMA samples from cystic fibrosis-affected individuals. The new approach greatly simplifies primer design, significantly increases the PCR multiplexing level, and decreases the overall primer cost. In addition, this assay is more readily amenable to automation and is therefore suitable for high-throughput genetic diagnostics.