Big-Dye Automated Sequencing
Reaction Mixes
10 ul Reaction:
- 2 ul Big Dye Reaction Buffer
- 2 ul Big Dye Terminator
- 2 ul 10 uM primer
- 2 ul Qiagen miniprep (200-500 ng template)
PCR Cycle (for PE 9600 or equivalent):
95oC for 2 minutes
95oC for 30 seconds
50oC for 15 seconds
60oC for 4 minutes
==> 30 cycles
4oC hold
Note: Ramping time is very important; should not be slower
than 1 degree/second.
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