Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching |
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Authors: | Maruyama Tatsuo Shinohara Toshimitsu Hosogi Takuya Ichinose Hirofumi Kamiya Noriho Goto Masahiro |
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Institution: | Department of Applied Chemistry, Graduate School of Engineering and Center for Future Chemistry, Kyushu University, Fukuoka 812-8581, Japan. |
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Abstract: | 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 oligonucleotide. |
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Keywords: | DNA Guanine quenching Fluorescent dye Fluorescence resonance energy transfer Single nucleotide polymorphisms |
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