Sample Preparation
All templates must be purified to ensure quality of sequencing. The best method is to use an agarose gel to verify the quality and quantity of your template (after purification). Samples must meet the requirements below to guarantee a good outcome.
- Primer: 10 pmol/ul. Please submit at least 1ul/reaction. The core provides the common universal primers such as M13-forward, reverse, T7, T3, Sp6, etc.
- DNA template: please submit at least 5ul/reaction.
DNA Template
| Template size (including vector) | Recommended Concentration |
|---|---|
| 100-200bp | 0.5-1.5 ng/ul |
| 200-500bp | 1.5-5 ng/ul |
| 500-1000bp | 5-10 ng/ul |
| 1000-2000bp | 10-50 ng/ul |
| 2000-4000bp | 50-150 ng/ul |
| 4000-6000bp | 150-250 ng/ul |
| 6000bp-10kb | 250-400 ng/ul |
| >10kb | >400 ng/ul |
Submission
- Place templates and primers in individual 1.5ml tubes. For multiple reactions to be done on the same template, place enough sample in one tube. Do not pre-mix your sample with primer. Samples will be discarded three weeks after results are sent unless the users request to keep them.
- Drop off sample at ABBR R651 cabinet, Cullen Building 604B, Alkek Building N1301, or Neurosensory Building NB428 with service request sheet by 12 p.m. Sample drop off after 12 p.m will be stored and processed the next working day.
Results
Data will be sent by email on the next morning, including Saturday.
Data Interpretation and Troubleshooting
If any sequencing failed, the core will perform troubleshooting together with the customer and repeat the sequencing up to three times for free.