Eppendorf Mastercycler Gradient - Hot Start PCR Example

37
7 Programming
7.6.2 Hot Start PCR
The "Pause" command is used to interrupt the program. The most-recently entered temperature is
maintained. A text explaining the reason for the interruption may be entered as required. After confirmation by
pressing , the program is continued.
The "Hold" command enables all possible temperatures to be maintained until the program is continued by
pressing .
In the exampleof a Hot Start PCR listed below, the program is stopped folllowing initial denaturation in order to
add a reaction component. After the final elongation, the block is cooled down to room temperature.
Hot Start PCR
Temperature control of tubes Control tube
Temperature control of lid lid 105°
Nowait auto
1 Initial denaturation 94° 2 min
2 Pause for adding substance Pause Adding
3 Denaturation 94° 15 sec
4 Annealing 55° 15 sec
5 Elongation 72° 30 sec
6 Number of cycles go to 3 Rep 29
7 Final elongation 72° 2 min
8 Cooling and storage Hold 20° enter
End end