Summary
The invention relates to a method of a mass-spectrometric analysis of known mutation sites in the genome, such as single nucleotide polymorphisms (SNPs), using the method of restricted primer extension.
The invention consists of the use of primers with a photocleavable linker. The linker creates a gap in a DNA strand which is almost the same size as a natural DNA building block (nucleoside). The linker forms a bridge over a base pair without inhibiting hybridization or enzymatic extension. However, the linker allows the primers to be cleaved after extension in order to obtain short DNA fragments which can be more easily detected on the mass spectrometer.
摘要
本发明涉及一种利用限制性引物延伸法对
基因组中已知突变位点(如单核苷酸多态性(SNP))进行质谱分析的方法。
本发明包括使用带有光可裂解连接体的引物。连接体在 DNA 链上形成一个缺口,其大小与天然 DNA 构建块(核苷)几乎相同。连接体在碱基对上形成桥接,不会抑制杂交或酶延伸。不过,连接子允许引物在延伸后被裂解,以获得更容易在质谱仪上检测到的短 DNA 片段。