Table 2.
FRET Conditions for Detection of the Polymorphisms within Intron 1 of RET Proto-Oncogene[Note]
|
Melting Conditions after Amplification |
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| PolymorphismGenotyped | Probesa | TemperatureTransition(°C) | TransitionRate(°C/s) | Gains (F1:F2:F3) |
| IVS1-1370C→T | Anchor: 5′- YgCTCACTCAgCCACAgCCgAggC pSensor: 5′- TgCACCgTgCCCCTgCTTg X | From 50 to 95 | .40 | |
| IVS1-1463T→C | Anchor: 5′- TTTTTgggAACTACCTTCTTgTTTTTCATSensor: 5′- Y-ACTgTATATTATTTTCCACTCACCgA p | From 40 to 95 | .20 | 1:45:30 |
Note.— We used 50 cycles of PCR at an annealing temperature of 66°C.
a
Boldface italics denote the nucleotides that are sensitive for the wild-type or the polymorphic allele. “X” represents fluorescein, and “Y” represents LC-Red 640. Some probes have modifications (represented by “p”) in their 3′ that disable extension of Taq polymerase.