Pyrophosphorolysis-activatable oligonucleotides may facilitate detection of rare alleles, mutation scanning and analysis of chromatin structures
Pyrophosphorolysis-activatable oligonucleotides may facilitate detection of rare alleles, mutation scanning and analysis of chromatin structures
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DOI：
10.1093/nar/30.2.598
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发表时间：
2002-01-15
影响因子：
14.9
通讯作者：
Sommer, SS
中科院分区：
文献类型：
--
作者：
Liu, Q;Sommer, SS
焦磷酸解激活聚合（PAP）最初是为了增强等位基因特异性PCR的特异性而开发的，用于在存在大量过量的野生型等位基因的情况下检测已知突变。PAP的高特异性来源于焦磷酸解介导的焦磷酸解可活化寡核苷酸（P*）的活化随后是活化寡核苷酸的延伸的串联偶联。在此，我们证明了基因工程DNA聚合酶大大提高了PAP的效率，使其成为检测罕见突变的实用技术。我们还表明，P* 寡核苷酸具有新的和意想不到的性质，在整个至少16个3 '-末端核苷酸的错配的高灵敏度。因此，每当沿着寡核苷酸的长度沿着需要高特异性时，PAP构成了具有潜在效用的技术平台。
Pyrophosphorolysis-activated polymerization (PAP) was initially developed to enhance the specificity of allele-specific PCR for detection of known mutations in the presence of a great excess of wild-type allele. The high specificity of PAP derives from the serial coupling of pyrophosphorolysis-mediated activation of a pyrophosphorolysis-activatable oligonucleotide (P*) followed by extension of the activated oligonucleotide. Herein, we demonstrate that genetically engineered DNA polymerases greatly improve the efficiency of PAP, making it a practical technique for detection of rare mutations. We also show that P* oligonucleotides have the novel and unexpected property of high sensitivity to mismatches throughout at least the 16 3'-terminal nucleotides. Thus, PAP constitutes a technology platform of potential utility whenever high specificity is required along the length of an oligonucleotide.