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Welcome to the University of Virginia Biology Department's Genome Sequencing Core!

The Department of Biology at the University of Virginia has recently acquired a Roche 454 FLX Genome Sequencing System. This state-of-the-art instrument serves as the cornerstone of our growing Comparative Genomics core. We are pleased to offer a range of genome sequencing services to individuals within and outside of our academic community.



Process Overview:  

Please visit this link for an comprehensive description of the sequencing process

Briefly, the sample workflow is as follows:

  • Customers submit a DNA sample that meets all quality control benchmarks set forth by the Genomics Core
  • A library is created via shearing and then ligation of process-specific adaptor molecules to the customer's sample
  • This library undergoes a concentration titration in an oil-in-water emulsion PCR reaction (emPCR) to determine the optimum amount of template library needed to produce enough clonal DNA for sequencing
  • A production emPCR is carried out to amplify sufficient DNA copies for sequencing
  • The library is deposited onto a sequencing plate and the sequencing run is carried out
  • Run data is stored for XXX hrs on the Core clusters, during which time it can be downloaded via FTP. The Core can also provide a hard copy customer data at at-cost.

454 Sequencing Highlights:

  • 400-600 megabase (Mb) with greater-than 1 million high-quality reads per run
  • 99% accuracy at base 400, > 99% at preceding bases (q20 = 400)
  • Unidirectional sequencing pass 400 bases (average) with Titanium chemistry
  • Ability to process dsDNA from genomic, organelle, BAC, YAC P1
  • Multiplex Identifier Taqs (MIDs) allow for sample pooling after library construction to reduce user expense and maximize data output

Applications*:

  • Prokaryotic/eukaryotic whole genome sequencing
  • Resequencing/deep sequencing
  • SNP detection
  • Metagenomics
  • Gene discovery
  • ChIP-seq/Methylation/Epigenetics

Current Services Offered**:

  • Standard (shotgun) library preparation
  • 3 - 8kb Paired-end library preparation
  • Sequencing from any library source (e.g. transciptome, amplicon, etc.)

Cost of Service:

  • Service prices vary depending upon the user's affiliation (academic, non-profit and commercial)

*For a comprehensive list of applications, please visit Roche's website

**Breadth of services will expand concomitantly with Roche product release.

 
 
 
             © Copyright 2009-10                   UVA Biology Dept. Genome Sequencing Core, Rm. 052C Gilmer Hall, 485 McCormick Rd., Charlottesville, VA 22903  (434) 243-9657