•
Methods Useful in Genetics and Oncology
0.5
r"'0
0
-
- T'"" U.
"-"
"'0 I
-0.5
-1
40
-"- Wild Type
-
Heterozygous
- - - Homozygous Mutant
........ No DNA
\
\
\
50
60
Temperature (O C)
70
Fig. 2. Genotyping curves for hereditary hemochromatosis mutation H63D (C187G). The fluorescein probe was designed to be complementary to the mutant sequence. Quenching was provided by two complementary guanosines
0.5
r- "'0 0
-
- T'"" U.
- "9 -0.5
-1
-1.5
40
50
60
_ .. - Wild Type
-
Heterozygous F50Sdei
- - - Homozygous F50Sdei
• • •• Heterozygous FS08C
•• -
Heterozygous IS07dei
...... No DNA
70
Temperature (O C)
Fig. 3. Melting troughs for genotyping the cystic fibrosis-associated mutation F508del. Deletion
of codon 508 results in a 3-bp mismatch loop in the probe, destabilizing the probe-template
hybrid by 9°C. Quenching was provided by a single complementary guanosine