PAP: a technology platform

描述（由申请人提供）： 焦磷酸解活化聚合（PAP）为聚合提供了一种新的方法， 从核酸中检索多种类型的信息。的特殊 PAP特异性来源于无活性的双脱氧末端寡核苷酸 （P*）。P* 被激活的焦磷酸解的3？末端核苷酸， 随后通过DNA聚合延伸活化的寡核苷酸。 最初的研究建立了原理证明，最近的效率 PAP的基因工程聚合酶大大改善， 对双脱氧核苷酸的高亲和力。在此，我们建议证明 PAP是一种平台技术。将开发三种新方法 该平台：i）等位基因特异性检测 10 expl 6 - 10 exp 9正常等位基因（PAP-A）的存在：ii）基于微阵列的 重测序（PAP-R）和iii）体内染色质结构分析 （LM-PAP）。这些方法具有许多应用，包括： 微小残留病检测或突变负荷测量（PAP-A）; ii）易患以下疾病的序列变体的分子流行病学分析： 复杂疾病或快速分子诊断（PAP-R）;和iii）分析 染色质结构作为印记、X-失活和基因的功能 表达或甲基化水平的定量（LM-PAP）。

DESCRIPTION (provided by applicant): Pyrophosphorolysis activated polymerization (PAP) offers a novel approach for retrieving multiple types of information from nucleic acids. The exceptional specificity of PAP derives from an inactive dideoxy terminated oligonucleotide (P*). P* is activated by pyrophosphorolysis of the 3? terminal nucleotide, followed by extension of the activated oligonucleotide by DNA polymerization. The initial studies established proof of principle and recently the efficiency of PAP was improved greatly with genetically engineered polymerases that have a high affinity for dideoxy nucleotides. Herein, we propose to demonstrate that PAP is a platform technology. Three novel methods will be developed on the platform: i) allele-specific detection of a single nucleotide change in the presence of 10expl6-10exp9 normal alleles (PAP-A): ii) microarray-based resequencing (PAP-R) and iii) analysis of in vivo chromatin structure (LM-PAP). These methods have many applications including: i) ultra sensitive detection of minimal residual disease or measurement of mutation load (PAP-A); ii) molecular epidemiological analysis of sequence variants that predispose to complex disease or rapid molecular diagnosis (PAP-R); and iii) analysis of chromatin structure as a function of imprinting, X-inactivation and gene expression or quantitation of the level of methylation (LM-PAP).