1. Prepare your template
Please prepare your template using our checklist:
- Ensure that any kits you use to prepare samples specify that DNA sequencing is a downstream application.
- Submit DNA templates in either sterile water or 10mM Tris pH 8.0.
- Supply 10 uL of DNA per reaction. Only 1 uL of DNA is used per reaction but providing additional volume allows for re-runs if necessary. If sample volume is a concern, please discuss with the Lab Coordinator.
- Review the following table for recommended quantity measurements. Final concentration will be in units of ng/uL or ug/uL depending on DNA template.
| Template DNAs | Quantity per reaction |
|---|---|
| 100-200 bp | 1-3 ng |
| 200-500 bp | 3-10 ng |
| 500-1000 bp | 5-10 ng |
| 1000-2000 bp | 10-40 ng |
| more than 2000 bp | 25-50 ng |
| Single-stranded DNA | 25-50 ng |
| Double-stranded DNA | 150-300 ng |
| Cosmid, BAC | 0.5-1.0 µg |
| Bacterial genomic DNA | 0.5-1.0 µg |
Fragment Analysis
Fragment analysis plates should be fully prepared by the client and will be run as is.