Techne Prime Elite Satellite - Manual (Page 39)

39
APPENDIX
TEMPLATE PROGRAMS
To help with programming, a number of program templates are saved in Techne Workbench which are
available to edit or can be run directly without changes. Details of the program templates are given
below.
Instrument and program defaults
Parameter 2 Step Template 3 Step Template RT-PCR Template
Heated Lid 105ºC 105ºC 105ºC
Heated lid before program On On On
Sample cooling On On On
Pause before program Off Off Off
Initial denaturation 94ºC, 05m00s 94ºC, 05m00s 94ºC, 05m00s
Hot start Off Off Off
Final extension 72ºC, 05m00s 72ºC, 05m00s 72ºC, 07m00s
Final hold 10ºC 10ºC 10ºC
Thermal cycling conditions
2 Step Template 3 Step Template RT-PCR Template
Stage 1
Number of cycles
30 30 1
Step 1 94.0ºC, 00m30s 94.0ºC, 00m30s 45.0ºC, 40m00s
Step 2 60.0ºC, 01m00s 55.0ºC, 00m30s 94.0ºC, 05m00s
Step 3 72.0ºC, 00m30s
Stage 2
Number of cycles
40
Step 1 94.0ºC, 01m00s
Step 2 55.0ºC, 00m50s
Step 3 72.0ºC, 01m00s
The Ice Bucket and Ligation programs are simple temperature holds, with the former having an infinite
hold at 4ºC and the latter at 15ºC. In both of these programs the heated lid is switched off and the
initial denaturation, hot start and final extension functions disabled.
INCREMENTED/DECREMENTED TIME AND TEMPERATURE
The Techne Workbench software allows the user to either increment or decrement the time and/or
temperature of a step within a cycling stage. The hold time or temperature of the step will be either
increased or decreased by the programmed amount each cycle following the first cycle.
This feature is used with applications such as touchdown PCR where the annealing temperature is
gradually decreased during the cycling process and long PCR, where due to the size of the product,
long annealing/extension times are required and the extension time is increased by 15 to 20s per
cycle during the final stages of the reaction.