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° C | 30 sec. | 1 cycle |
| 96° C | 10 sec. | 25 cycles |
| 50° C | 5 sec. | |
| 60° C | 4 min. | |
| 4° C | hold | |
| GC Rich Thermal Cycling Conditions | ||
|---|---|---|
| 96° C | 1 min. | 1 cycle |
| 98° C | 15 sec. | 30 cycles |
| 55° C | 1 sec. | |
| 62° C | 4 min. | |
| 60° C | 2 min. | 1 cycle |
| 4° C | hold | |
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