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High-throughput Sequencing

 

High-throughput Sequencing provides an array of services for Sanger sequencing projects that have fifty or more 96-well plates in a two month period.  Customers are allowed to submit samples starting at any stage in the sequencing process, from glycerol stocks for culturing to plates that are ready to load on the sequencers.  Conditions for each project are optimized to produce the best sequencing data and automation is used where possible.  The Qiagen 9600 BioRobot is used for plasmid preps, while the Robbins Hydra is used for precise reaction set-up.  Thermocycling conditions are optimized on the BioRad/MJ Research DNA tetrads.  BigDye terminator chemistry is used for Sanger sequencing reactions and all sequencing is performed on the AB 3730xl DNA Analyzer.  All 384- and 96-well format plates are labeled with a barcode, and a high-throughput laboratory information management system (HTLIMS) is used to track sample flow from plate submission through sequencing.

No more than 2 primers are allowed per project for HT Full Service and HT No Prep projects.  Those projects that do not meet the minimum plate requirements, or those with more than 2 primers, are considered medium-throughput projects and must be submitted through the CoreLIMS website (unicorn.biotec.illinois.edu).

 

HT  Plasmid

 

  • User submits purified PCR products or plasmids.
  • Service includes sequencing reaction, ethanol precipitation clean-up, electrophoresis on the AB 3730xl and data pickup site.
  • $285.00 per 96-well plate for first primer, same price for second primer if desired.

 

HT BAC-end Sequencing

  • User submits purified BACs.
  • Service includes sequencing reaction, ethanol precipitation clean-up, electrophoresis on the AB 3730xl and data pickup site.
  • $285.00 per 96-well plate, same price for second primer if desired.

 

HT Ready-to-Load With Clean-up

  • User submits unpurified BigDye reactions of PCR products, plasmids or BACs.
  • Service includes column purification, electrophoresis on the AB 3730xl and data pickup site.
  • $148.00 per 96-well plate.

 

HT Ready-to-Load

  • User submits purified BigDye reactions of PCR products, plasmids or BACs.
  • Service includes electrophoresis on the AB 3730xl and data pickup site.
  • $67.00 per 96-well plate

 

Customers are charged for a full plate even if some wells are left empty.  If the percentage of successful reactions on a plate is below 80% (defined as 200 high quality bases) due to a problem deemed to be a technical issue in the Core Facility, the price per plate will be adjusted proportionally downward.  Sequencing success rates will be calculated for each plate and those with success rates of 30% and above will be billed.  The investigator will be billed based on the combined success rates of the original and the repeated plates. 

 

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.  For further information please contact the Core staff at dna-seq@illinois.edu or 217-333-9520 to discuss ways we can be of assistance in achieving your project goals or to receive a quote for your project.  

Facility users other than State of Illinois institutions are subject to a 28.6% indirect cost rate for federally funded projects. For off-campus projects a contract must be negotiated through University of Illinois Grants and Contracts.  For further information contact Dr. Mark A. Mikel (217-244-0144; mmikel@illinois.edu ), Associate Director of the Roy J. Carver Biotechnology Center.