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首页> 外文期刊>Journal of mass spectrometry: JMS >Nucleic acid and SNP detection via template-directed native chemical ligation and inductively coupled plasma mass spectrometry
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Nucleic acid and SNP detection via template-directed native chemical ligation and inductively coupled plasma mass spectrometry

机译：通过模板定向的天然化学连接和电感耦合等离子体质谱法检测核酸和SNP检测

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摘要

Detection of nucleic acids and single nucleotide polymorphisms (SNPs) is of pivotal importance in biology and medicine. Given that the biological effect of SNPs often is enhanced in combination with other SNPs, multiplexed SNP detection is desirable. We show proof of concept of the multiplexed detection of SNPs based on the template-directed native chemical ligation (NCL) of PNA-probes carrying a metal tag allowing detection using ICP-MS. For the detection of ssDNA oligonucleotides (30 bases), two probes, one carrying the metal tag and a second one carrying biotin for purification, are covalently ligated. The methodological limit of detection is of 29 pM with RSD of 6.7% at 50 pM (n = 5). Detection of SNPs is performed with the combination of two sets of reporter probes. The first probe set targets the SNP, and its yield is compared with a second set of probes targeting a neighboring sequence. The assay was used to simultaneously differentiate between alleles of three SNPs at 5-nM concentration.
机译：检测核酸和单核苷酸多态性（SNPs）在生物学和医学中具有关键重要性。鉴于SNP的生物学效果通常与其他SNP结合增强，期望多路复用的SNP检测。基于携带使用ICP-MS的金属标签的PAN-探针的模板定向的本机化学连接（NCL），显示了允许使用ICP-MS检测的模板导向的本机化学连接（NCL）的模板定向检测概念证据。为了检测SSDNA寡核苷酸（30个碱基），两种探针，携带金属标签的探针和携带生物素的第二个纯化，是共价连接。检测的方法论极限为29pm，50 pm的RSD为6.7％（n = 5）。通过两组报道探针的组合进行SNP的检测。第一探针组靶向SNP，将其产率与靶向相邻序列的第二组探针进行比较。用于在5nm浓度下同时区分三个SNP的等位基因。

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