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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