Combination of a modified block PCR and endonuclease IV-based signal amplification system for ultra-sensitive detection of low-abundance point mutations

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• 作者：Xianjin Xiao ; Anqin Xu ; Junqiu Zhai ; Meiping Zhao
• 关键词：PCR ; polymerase chain reaction ; dNTP ; deoxyribonucleoside triphosphate ; UDG ; Uracil-DNA Glycosylase ; Endo IV ; endonuclease IV ; 位exo ; lambda exonuclease ; HRM ; high resolution melting ; LOD ; limit of detection ; AS-PCR ; allele specific PCR ; COLD-PCR ; co-amplification at lower denaturation temperature-PCR ; WT ; wild type ; MT ; mutant ; AP ; apurinic/apyrimidinic
• 刊名：Methods
• 出版年：15 December, 2013
• 年：2013
• 卷：64
• 期：3
• 页码：255-259
• 全文大小：843 K

文摘

By combination of a modified block PCR and endonuclease IV-based signal amplification system, we have developed a novel approach for ultra-sensitive detection of point mutations. The method can effectively identify mutant target sequence immersed in a large background of wild-type sequences with abundance down to 0.03% (for C 鈫?#xA0;A) and 0.005% (for C 鈫?#xA0;G). This sensitivity is among the highest in comparison with other existing approaches and the operating procedures are simple and time saving. The method holds great potential for future application in clinical diagnosis and biomedical research.