DNA Core / Sanger Facility
Core / Sanger Facility
Please visit our CoreLims website (https://corelims.biotech.illinois.edu) to submit your fragment analysis orders and Sanger sequencing orders.
Sample drop boxes are available at 4 campus locations
- To use the drop boxes, fill out the online form as usual and put it into a sealed bag with your samples and primers.
- Samples in the drop boxes will be picked up at 2:00pm, Monday through Friday.
- Results for these samples and any received at the sequencing lab by 5:00pm will be posted by 5:00pm the following business day.
- Everitt Laboratory
1406 West Green Street, Urbana- Located near the receiving area
- Chemical and Life Sciences Lab (MCB), Room C107
601 South Goodwin Avenue, Urbana- Located directly outside of the CLSL Life Sciences Storeroom
- IGB Building, Room 1201
1206 West Gregory Avenue, Urbana- Located directly inside of the mail room/central receiving.
- Veterinary Medicine Basic Sciences Building, Room 1828
2002 South Lincoln Avenue, Urbana- Located directly inside of the mail room.
Please contact us if you have any questions about using the drop boxes. 217-265-6814 or dna-seq@illinois.edu
Sanger Sequencing Overview
For low- and high-throughput Sanger DNA sequencing, we accept purified PCR products, plasmids and large DNA constructs such as cosmids, fosmids and BACS. Low-throughput orders are defined as 1-95 sequencing reactions submitted at any one time. High-throughput orders are defined as 1 or more plates of sequencing reactions submitted at any one time. Samples must be submitted in 96-well plate format to receive the high-throughput rate reduction.
We offer three types of sequencing options (see below) and our typical turn-around time is one business day from the date of submission. All orders are completed on a first-come, first-served basis, regardless of affiliation. Please see the Sample Submission Guidelines on the CoreLims website for information about sample concentration, primers, and submitting samples.
Troubleshooting is free and we encourage customers to contact us if they are having difficulties with their sequencing project. We offer a Troubleshooting Manual on our CoreLims website to assist customers.
Sequencing option
- User submits purified PCR products, plasmids, cosmids, fosmids or BACs.
- User submits custom primers or uses those provided by the Core facility.
- Service includes sequencing reaction, column purification and electrophoresis on the AB 3730xl.
Ready-to-Load with Clean-Up option
- User submits unpurified BigDye reaction.
- Service includes column purification and electrophoresis on the AB 3730xl.
Ready-to-Load Option
- User submits purified BigDye reaction.
- Service includes only electrophersis on the AB 3730xl.
- This service is for 96-well plates only. You may not submit samples in individual tubes.
Primer Walking
- If your DNA region of interest is longer than 800 bp, you may want to do a primer walk to sequence the entire region.
- We edit the sequence, design and order primers, do the sequencing reactions, then align all the sequences in Sequencher.
- We send you the clean, full length sequence of your region of interest.
- $10.00 per designed primer plus the cost of the primer and the sequencing reaction.
Fragment Analysis Overview
For Fragment Analysis, we accept unpurified (unless otherwise directed) PCR products generated by microsatellite, AFLP, RFLP, TRFLP, ARISA or other protocols. Our capillary instruments can analyze fluorescently-labeled fragments up to 1200bp in length, and they provide high run-to-run consistency with low background noise. For sample analysis, we provide our customers with free access to a shared copy of the GeneMapper software via remote connection to a computer at the Core Facility. Our typical turn-around time is one business day from the date of submission, and all orders are completed on a first-come, first-served basis, regardless of affiliation.
Size Standards
Several size standards are available to analyze fragments with 3-dye or 4-dye multiplexing. Size standards are provided by our facility and do not need to be purchased separately. Please contact us if you would like to use a custom size standard.
| Size Standard | Size Range (bp) | Blue Dye Label | Green Dye Label | Yellow Dye Label | Red Dye Label |
| LIZ120 | 50-120 | dR110 | dR6G | dTAMRA | dROX |
| ROX400 | 50-400 | 6-FAM | HEX | NED | – |
| ROX500 | 50-500 | 6-FAM | HEX | NED | – |
| LIZ500 | 50-500 | 6-FAM | VIC | NED | PET |
| LIZ600 | 50-600 | 6-FAM | VIC | NED | PET |
| ROX1000 | 50-1000 | 6-FAM | HEX | NED | – |
| LIZ1200 | 35-1200 | 6-FAM | VIC | NED | PET |
Submission Information
We provide one free 96-well test plate of serial dilutions to determine optimal DNA concentrations for each new customer. Please see the Sample Submission Guidelines on the CoreLims website for information about choosing size standards and primer dye labels, making a dilution plate, submitting orders and accessing analysis software. Due to the limitations of our capillary electrophoresis platform, we cannot successfully perform temperature-dependent analyses such as DGGE or SSCP.
Contact
DNA Services
Director: Alvaro G. Hernandez Ph. D.
University of Illinois at Urbana-Champaign
Roy J. Carver Biotechnology Center
1201 W. Gregory Drive
329 Edward R. Madigan Laboratory (ERML)
Urbana, IL 61801
(217) 244-3480
