Reaction Conditions

Sequencing Reaction Setup Components Volume

Template DNA* *either 300-500 ng plasmid or 10-20 ng PCR product for every 200 bp sequence
Primer 4pmol
Big Dye Terminator V3.1 8 ul
Total Volume 20 ul

Standard Thermal Cycling Conditions
96 deg C 30 sec 1 cycle
96 deg C 10 sec 25 cycles
50 deg C 5 sec
60 deg C 4 min
4 deg C hold  

GC rich Thermal Cycling Conditions
96 deg C 1 min 1 cycle
98 deg C 15 sec 30 cycles
55 deg C 1 sec
62 deg C 4 min
60 deg C 2 min 1 cycle
4 deg C hold  
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