TY - JOUR
T1 - Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching
AU - Maruyama, Tatsuo
AU - Shinohara, Toshimitsu
AU - Hosogi, Takuya
AU - Ichinose, Hirofumi
AU - Kamiya, Noriho
AU - Goto, Masahiro
N1 - Funding Information:
This research was supported by the GOTO Project of the “Science and Technology Incubation Program in Advanced Region” of Innovation Plaza Fukuoka under the Japan Science and Technology Agency and by a Grant-in-Aid for the 21st Century COE Program, “Functional Innovation of Molecular Informatics,” from the Ministry of Education, Culture, Science, Sports, and Technology of Japan.
PY - 2006/7/1
Y1 - 2006/7/1
N2 - 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.
AB - 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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U2 - 10.1016/j.ab.2006.03.056
DO - 10.1016/j.ab.2006.03.056
M3 - Article
C2 - 16701075
AN - SCOPUS:33744531012
VL - 354
SP - 8
EP - 14
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -
