<aside> 📂

Unlisted protocol. This exists in the repo but was never added to the protocols index, so it has not been visible on the lab docs site.

</aside>

In addition to sequencing technologies available at the Biotech Center on campus, full-length plasmid and amplicon sequencing is available through Plasmidsaurus.

UW Biotech Center sequencing

<aside> 🔑

An account with the UWBC Requester Portal is required to submit any samples to campus sequencing services. Access can be requested by emailing [email protected] or by following the instructions here.

</aside>

The UW Biotech Center's Interactive Submission Guideline provides application-specific instructions for all sample types. Follow these sample preparation, naming, labeling, and submission guidelines to properly submit samples for long-read plasmid or amplicon sequencing services.

Plasmidsaurus sequencing

Materials

• 8-well strip tubes
• Parafilm
• 50 mL conical tube
• Plasmids or amplicons (>10 μL at 30 ng/μL)

Protocol

1. Use the Qubit to measure the concentration of each plasmid or amplicon and normalize each to 30 ng/μL with nuclease-free water. Add at least 10 μL of each plasmid to a tube in an 8-well strip tube. Label the samples by writing the order label on the first tube and the strip number on the second tube of each strip — no additional labeling is necessary. See the Plasmidsaurus guidelines for any clarifications.
2. When all samples are ready and aliquoted into the strip tubes, use parafilm to seal the caps so they do not open during transit. Place the strip tubes in a 50 mL conical tube, and add a copy of the sample submission form and your samples to a Ziploc bag.
3. Submit a plasmid sequencing order through Plasmidsaurus. You will need to create an account if you do not already have one. Enter the plasmid names in the exact order they were deposited into the 8-well strip tubes.
4. Time your sample drop-off so that the samples are not sitting out at room temperature for too long. Include a copy of the submission form with your sample, put an additional physical copy on Kathy's desk, and put the digital receipt in the P-card receipts folder on Box.
5. Plasmidsaurus will email the results, usually within 24 hours. Transfer the results to Box > Data > Plasmidsaurus, in a folder labeled with the submission date.

Campus dropbox locations

<aside> 📍

Needs update. The source describes dropboxes "located throughout campus" but the table lists only one. Add the others, or reword.

</aside>