Techne Prime Elite Satellite Uživatelský manuál stáhnout pdf (Strana 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

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

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.

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