Applied Biosystems SimpliAmp Thermal Cycler - Page 73

Basic PCR Taq 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 95.0 72.0
Hold Time 60 15 420
Step 2 RampRate — 6.0 6.0 —
Temperature 55.0 4.0
Hold Time 15 Infinite
Step 3 RampRate — 6.0 — —
Temperature 72.0
Hold Time 30
BigDye
™
Direct Cycle Sequence Method
Cycle/Step
Stage Reaction
volume
(µL)
Cover
temp. (°C)
1 2 3 4 5
Cycles 1 1 1 25 1 50 105.0
Step
1
RampRat
e
4.0 6.0 6.0 6.0 4.0
Temperat
ure
37.0 80.0 96.0 96.0 4.0
Hold Time 900 120 60 10 Infinite
Step
2
RampRat
e
— — — 6.0 —
Temperat
ure
50.0
Hold Time 5
Step
3
RampRat
e
— — — 6.0 —
Temperat
ure
60.0
Hold Time 75
Appendix E Predefined run methods
Location of power point and ports on the instrument
E
SimpliAmp
™
Thermal Cycler User Guide 73