n Applications in Genetics
a. Wnd type probe
b. Mutant probe
us
52 ~ 56 58 60 62 64 66 68 10 12 14 76
52 ~ 56 58 60 62 64 66 68 70 12 74 76
Temp erature ("C)
- - Normal tissue
- - tumor
-noDNA
Temperature ("C)
I Wid type peak
I Mutant peak
Fig. 3. Detection of the BRCAI 3519G~T mutation in micro dissected tumor and normal tissue
from a carrier of this mutation. As expected, normal tissue showed two melting peaks and is heterozygote for the mutation. The tumor tissue, however, shows only the mutant melting peak at
60.2 (wild type probe) or 63.7 °C (mutant probe).
References
1. Pals G, Young C, Mao HS, Worsham MJ (2001) Detection of a single base substitution in a single cell using the LightCycler. J Biochem Biophys Methods 47:121-129
2. Peelen T, van Vliet M, Petrij-Bosch A et al. (1997) A high proportion of novel mutations in
BRCAI with strong founder effects among Dutch and Belgian hereditary breast and ovarian
cancer families. Am J Hum Genet 60: 1041-1 049
3. http://www.nhgri.nih.gov/lntramural_research/Lab _transfer/Bic!
4. Pals G, Pindolia K, Worsham MJ (1999) A rapid and sensitive approach to mutation detection using real-time polymerase chain reaction and melting curve analyses, using BRCAI as
an example. Mol Diagn 4:241-246
a. Wnd type probe
b. Mutant probe
us
52 ~ 56 58 60 62 64 66 68 10 12 14 76
52 ~ 56 58 60 62 64 66 68 70 12 74 76
Temp erature ("C)
- - Normal tissue
- - tumor
-noDNA
Temperature ("C)
I Wid type peak
I Mutant peak
Fig. 3. Detection of the BRCAI 3519G~T mutation in micro dissected tumor and normal tissue
from a carrier of this mutation. As expected, normal tissue showed two melting peaks and is heterozygote for the mutation. The tumor tissue, however, shows only the mutant melting peak at
60.2 (wild type probe) or 63.7 °C (mutant probe).
References
1. Pals G, Young C, Mao HS, Worsham MJ (2001) Detection of a single base substitution in a single cell using the LightCycler. J Biochem Biophys Methods 47:121-129
2. Peelen T, van Vliet M, Petrij-Bosch A et al. (1997) A high proportion of novel mutations in
BRCAI with strong founder effects among Dutch and Belgian hereditary breast and ovarian
cancer families. Am J Hum Genet 60: 1041-1 049
3. http://www.nhgri.nih.gov/lntramural_research/Lab _transfer/Bic!
4. Pals G, Pindolia K, Worsham MJ (1999) A rapid and sensitive approach to mutation detection using real-time polymerase chain reaction and melting curve analyses, using BRCAI as
an example. Mol Diagn 4:241-246