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.
| Original language | English |
|---|---|
| Pages (from-to) | 8-14 |
| Number of pages | 7 |
| Journal | Analytical Biochemistry |
| Volume | 354 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jul 1 2006 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching. / Maruyama, Tatsuo; Shinohara, Toshimitsu; Hosogi, Takuya; Ichinose, Hirofumi; Kamiya, Noriho; Goto, Masahiro.
In: Analytical Biochemistry, Vol. 354, No. 1, 01.07.2006, p. 8-14.Research output: Contribution to journal › Article
}
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
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.
UR - http://www.scopus.com/inward/record.url?scp=33744531012&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33744531012&partnerID=8YFLogxK
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 -