Big-Dye Automated Sequencing

Reaction Mixes

10 ul Reaction:

  • 4 ul Big Dye Mix
  • 1 ul 4 pM primer
  • 5 ul Qiagen miniprep (200-500 ng template)


PCR Cycle (for PE 9600 or equivalent):

95oC for 5 minutes

95oC for 10 seconds
50oC for 5 seconds
60oC for 4 minutes
  ==> 30 cycles
4oC hold

Note: Ramping time is very important; should not be slower than 1 degree/second.