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Core / Sanger Facility

Please visit our CoreLims website (https://unicorn.biotec.illinois.edu) to submit your fragment analysis orders and Sanger sequencing orders.

Leslie D. Benson
Leslie D. Benson

Lab Supervisor

The Core Facility provides Sanger DNA sequencing and fragment analysis services to researchers at domestic and international academic institutions, government agencies and private companies. All sequencing and fragment analysis samples are analyzed on two Applied Biosystems 3730xl DNA Analyzers to provide customers with the most sophisticated capillary sequencing technology to date.  These automated sequencers run with 50 cm capillary arrays and are extremely fast and accurate. We offer two levels of service options, based on the number of samples in your project and the way you submit them. 

  • Low-throughput:  1-95 samples submitted in tubes.
  • High-throughput:  1 or more 96-well plates submitted at any one time.

Our CoreLims website (unicorn.biotec.illinois.edu) is used to submit and track orders, retrieve data, gain access to analysis software and obtain troubleshooting assistance.  

 

Sanger Sequencing

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.

Sequence Editing

  • Sequences are edited up to 800 bp when possible, or the end of the PCR product if it is shorter than 800 bp.
  • Ambiguous or miscalled bases are corrected manually. 
  • $10.00 per sample.

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 services

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.

Pricing Information

Core Sequencing

Price

Sanger - High-Throughput Ready to Load (96-well Plate) - no clean up

$64

Sanger - High-Throughput Ready to Load (96-well Plate) - clean up

$148

Sanger - High-Throughput Sequencing (96-well plate)

$390

Sanger - Low-Throughput Sequencing (One Sample)

$4.90

Sanger - Low-Throughput Ready to Load (One Sample, includes clean-up)

$2.10

Fragment Analysis - High-Throughput (96-well Plate)

$85

Fragment Analysis - Low-Throughput (One Sample)

$2.70

Customers are encouraged to discuss any problems or concerns with facility personnel, as our goal is to provide only the highest-quality sequencing data and support. For consultations, please contact the Core staff at dna-seq@illinois.edu or 217-265-6814.

All work performed by the Roy J. Carver Biotechnology Center (CBC) should be acknowledged in scholarly publications, posters, and presentations. Proper recognition allows us to measure the impact of our work and supports our initiatives in obtaining sponsored funding. In addition, any CBC personnel who make a substantial intellectual or experimental contribution are deserving of further recognition as co-author.

This is an abridged list of our service prices and subject to change at any time. If you are an off-campus customer, please complete the Non-UI user form and submit with your first order.