The invention presents a method for examining genetic material (deoxyribonucleic acid, DNA) to detect the presence of pre-known mutations, especially single nucleotide polymorphisms (SNP), using mass spectrometry with ionization by matrix-assisted laser desorption (MALDI). The invention uses nucleoside triphosphates with modified sites for the method of primer extension in a duplicating, enzymatic reaction and at least partially removal of primers from the extension product, in combination with product neutralization by chemical treatment of the modified sites, so that the resulting DNA products can be, by using special matrix materials, preferredly ionized in an adduct-free form over other constituents in the reaction solution without any further cleaning. The method is particularly suitable for simultaneous identification of several mutations by multiplexing.
本发明提出了一种利用基质辅助激光解吸(
MALDI)电离质谱法检测遗传物质(
脱氧核糖核酸,DNA)以发现是否存在已知突变,特别是单
核苷酸多态性(SNP)的方法。本发明使用具有修饰位点的核苷
三磷酸酯,在复制、酶促反应中进行引物延伸,并至少部分去除延伸产物中的引物,同时通过
化学处理修饰位点对产物进行中和,这样,通过使用特殊的基质材料,所得到的 DNA 产物最好能以不含加合物的形式电离出反应溶液中的其他成分,而无需进一步清洗。该方法尤其适用于通过多路复用同时鉴定多个突变。