From GenoSeq
Contents |
Sequencing Customer Information Page
Services
The facility offers two sequencing services:
- Full Service, in which we carry out the sequencing reactions on your samples for you.
- Ready-to-Run, if you prefer to do the sequencing reaction yourself and bring in the reacted sample.
Full Service
| Sample Type | Volume/Sequence | DNA conc. |
|---|---|---|
| Plasmid | 8 µl | 200 ng/µl |
| PCR product | 8 µl | 20 ng/µl or an aliquot of your cleaned PCR product |
| BAC | 11 µl | 230 ng/µl |
Sample and primer names are limited to 6 characters with no spaces.
Plasmids submitted should contain at least one commonly used sequencing primer site (M13F, M13R, T7, T3, Sp6, and BGHR.) The Facility provides these common primers. If your plasmid doesn't have one of these primer sites, you must provide your own specific primers. If you want to use your own primer we need 5.0 µl per reaction at a concentration of 5-10 µM (5-10 pmol/µl).
Sequences for Primers provided by the Core
M13F primer: -21 M13F : TGT AAA ACG ACG GCC AGT
M13 R primer: -28 M13R : AGG AAA CAG CTA TGA CCA T
T7 primer: TAA TAC GAC TCA CTA TAG GG
T3 primer: ATT AAC CCT CAC TAA AGG GA
SP6 primer: ATT TAG GTG ACA CTA TAG
BGHR primer: TAG AAG GCA CAG TCG AGG
Ready-To-Run
You do the sequencing reaction yourself and we run the samples. Bring us your wet DNA samples after column purification. If you use ethanol precipitation to purify your sample you must resuspend it in 20µl of distilled water. Alternatively, we can clean the samples for you for a flat $20 charge per daily submission regards of the number of samples up to 96. You may bring your sample in micro-centrifuge tubes or on a PCR plate if you have a large number of samples that need cleaning.
When you submit Ready-to-Run samples, the Core does not process your sample in any way. We merely pipette the sample from your tube onto an optical plate, and then put the plate on the capillary sequencer. We run regularly scheduled validation checks of all our instruments with control samples to verify that all equipment is functioning properly. Therefore, if the run is successful for the other samples on the plate, and your sample fails, the Core does not accept responsibility for the failure. You will be charged for running both the failed sample and any reruns. We will be happy to help you troubleshoot your sequencing reactions, or you can use our Full-Service option, and let us do the work for you.
Sample Handling at the Facility
- All samples are entered into a database that gives each entry an ID number for customer results and data storage purposes.
- Plasmid samples and PCR products are quantitated by running them on an agarose gel next to a DNA standard before sequencing reactions are performed.
- Remaining PCR products, extracted plasmids, and specific primers will be returned to customers upon request.
Sequencing Reactions
All sequencing is performed on a Applied Biosystems 3730 Capillary DNA Analyzers.
Turn-around Time
The turn-around time is 1-2 working days for Ready-to-Run samples and 1-2 working days for Full Service samples. Occasional delays may occur if we experience unusually heavy loads or equipment failure.
Results
Results will be available as computer files. Results can be retrieved either directly from the facility or from Web-Seq, our interactive web-based sequence retrieval system. For full instructions on how to use Web-Seq, go to the Web-Seq site (http://www.genetics.ucla.edu/webseq/) and click on the "help" button.
Technical Advice
We have staff that can assist you in interpreting your results and trouble-shooting problems.
Data Storage
All data is backed up daily. Raw data is stored for 1 month. Analyzed data is permanently archived and backed up off-site.
Cost
Costs are $12.00 for Full Service reactions and $5.00 for Ready-to-Run reactions.
Also available at the core:
| BigDye Terminator | 200 µl |
| Sequencing Buffer | 400 µl |
| sequence cleanup columns | 50 pkg. |
An account will be set up for each P.I. using the facility. Customers wishing to set up a new account should bring a UCLA Recharge Order Request (form P-39) or equivalent billing information.
Facility Hours
- Monday through Friday: 9 AM to 5 PM
- Saturday and Sunday: closed
Location
- 5309 Gonda Center on the UCLA campus.
- 695 Charles Young Drive South
- Los Angeles, CA 90095
- E5 on the South Campus sector of the UCLA Campus Map
- Open 9AM to 5PM Mon-Fri, excluding university holidays
Contact
- Uma Dandekar, GenoSeq Core Assistant Director
- Email: uma@ucla.edu
- Phone: 310-267-2461
- Fax: 310-794-5446