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Brilliant Core Reagent Kits for QPCR and QRT-PCR

2; 200 M each of dGTP, dATP, dCTP, and 400 M of dUTP; 0.5 units of UNG; optimal concentrations of sequence-specific primers and probe; and 2.5 U of SureStart Taq DNA polymerase in a final volume of 50 l. A no-template control was included. The PCR cycling conditions for molecular beacons are as follows: 50C for 2 minutes; 95C for 30 seconds; 55C for 1 minute; and 72C for 30 seconds. The thermal cycling conditions were 50C for 2 minutes; 95C for 10 minutes; for monitoring TaqMan reactions, this was followed by 40 cycles of 95C for 15 seconds and 60C for 1 minute. A temperature cycler designed to detect changes in fluorescence in real time was used.

REFERENCES

  1. Higuchi, R., et al. (1992) Bio/Technology 10: 413-417.

  2. Higuchi, R., et al. (1993) Bio/Technology 11: 1026-1030.

  3. Whitcombe, D. et al (1999) Nature Biotechnol. 17: 804-807.

  4. Nielson, K. et al (1993) Strategies 6: 45-46


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