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Table 2 Fast COLD-PCR Cycling Program

From: COLD-PCR enhanced melting curve analysis improves diagnostic accuracy for KRAS mutations in colorectal carcinoma

Step Temp Time Analysis Mode Acquisition Mode
UNG 40°C 10 m None None
Denaturation 94°C 10 m None None
Regular-PCR (10 cycles) 94°C 1 s None None
  52°C 20 s None None
  72°C 10 s None None
COLD-PCR (40 cycles) 81 °C ( T c ) 1 s None None
Ramp 2.0°C/s 52°C 20 s Quant Single
  72°C 10 s None None
Melting curve 94°C 5 s None None
  45°C 30 s None None
  80°C Ramp 0.15°C/s Melting Continuous
Cooling 40°C 10 s None None
  1. UNG: Uracil-N-Glycosylase; Temp: Temperature.