BigDye
™
Direct PCR Run Method
Cycle/Step
Stage Reaction
volume
(µL)
Cover
temp. (°C)
1 2 3 4
Cycles 1 35 1 — 50 105.0
Step 1 RampRate 6.0 6.0 6.0 —
Temperature 95.0 96.0 72.0
Hold Time 600 3 120
Step 2 RampRate — 6.0 6.0 —
Temperature 62.0 4.0
Hold Time 15 Infinite
Step 3 RampRate — 6.0 — —
Temperature 68.0
Hold Time 30
BigDye
™
Kit Fast Run Method
Cycle/Step
Stage Reaction
volume
(µL)
Cover
temp. (°C)
1 2 3 4
Cycles 1 25 1 — 50 105.0
Step 1 RampRate 6.0 6.0 6.0 —
Temperature 96.0 96.0 4.0
Hold Time 60 10 Infinite
Step 2 RampRate — 6.0 — —
Temperature 50.0
Hold Time 5
Step 3 RampRate — 6.0 — —
Temperature 60.0
Hold Time 75
Appendix E Predefined run methods
Location of power point and ports on the instrument
E
74
SimpliAmp
™
Thermal Cycler User Guide