Sequence Reaction Guidelines
1/16th Big Dye Reaction in 5ul
0.5ul of Big Dye Mix
0.75ul of Applied Biosystems 5x dilution buffer (provided with kits)
3-5pmols of primer
template per the following guidelines
q.s to 5ul
| Template | Quantity Per Reaction |
|---|---|
| 100–200 bp PCR product | 2-4 ng |
| 200–500 bp PCR product | 0.5-3 ng |
| 500–1000 bp PCR product | 2-20 ng |
| 1000–2000 bp PCR product | 5-40 ng |
| >2000 bp PCR product | 10-50 ng |
| Single-stranded plasmid: | 10-50 ng |
| Double-stranded plasmid: | 50-300 ng |
| Cosmid, BAC | 0.5–1.0 µg |
| Bacterial genomic DNA | 2–3 µg |
Cycling Conditions
For 1 cycle:
96° 1min
For 25 cycles:
96° 10sec
50° 5sec
60° 4min
For 1 cycle:
4° ¥
Reactions should be submitted in PCR strip tubes with strip caps. Tubes with individual or attached caps will not be accepted. We recommend strip tubes from Genesee Scientific, P/N 22-705.