Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching
Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching
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DOI：
10.1016/j.ab.2006.03.056
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发表时间：
2006-07-01
影响因子：
2.9
通讯作者：
Goto, Masahiro
中科院分区：
文献类型：
--
作者：
Maruyama, Tatsuo;Shinohara, Toshimitsu;Goto, Masahiro
荧光标记的DNA探针的鸟嘌呤淬灭是用于检测DNA链的靶位点中的突变的有力工具。然而，与靶位点相邻的不同鸟嘌呤可干扰点突变的检测，导致不令人满意的灵敏度。在目前的研究中，我们开发了一种简单的方法来提高灵敏度的鸟嘌呤淬灭法使用掩蔽DNA寡核苷酸。发现简单添加掩蔽DNA寡核苷酸掩蔽靶寡核苷酸中不同鸟嘌呤对荧光的干扰，并增强野生型和突变型寡核苷酸之间淬灭比的差异。基于这一策略，我们成功地区分各种突变的野生型YMDD基序的B型肝炎病毒DNA聚合酶基因使用鸟嘌呤淬灭与掩蔽寡核苷酸。(c)2006年爱思唯尔公司All rights reserved.
Guanine quenching of a fluorescence-labeled DNA probe is a powerful tool for detecting a mutation in a targeted site of a DNA strand. However, a different guanine adjacent to a targeted site can interfere with detection of a point mutation, resulting in unsatisfactory sensitivity. In the current study, we developed a simple method to improve sensitivity of the guanine quenching method using a masking DNA oligonucleotide. The simple addition of a masking DNA oligonucleotide was found to mask the interference of a different guanine in a target oligonucleotide on fluorescence and to enhance difference in the quenching ratio between wild-type and mutant oligonucleotides. Based on this strategy, we succeeded in discriminating various mutations from the wild-type YMDD motif of the hepatitis B virus DNA polymerase gene using guanine quenching with a masking oligonucteotide. (c) 2006 Elsevier Inc. All rights reserved.