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The UT Genomics Core (UTGC), along with the Center for Biotechnology (CEB) and the Office of Research, Innovation, and Economic Development (ORIED), are pleased to announce the arrival of the Illumina NovaSeq high throughput sequencing system.
The UT Genomics Core provides high-throughput next-generation sequencing services to all interested on-campus and off-campus researchers. Sequencing is performed using the Illumina NovaSeq, MiSeq, or Oxford Nanopore MinION instruments located in room 407 of the Science and Engineering Research Facility (SERF) within the Center for Environmental Biotechnology (CEB). We recommend consultation with UTGC staff prior to project initiation. While we strive for successful sequencing every time, we cannot guarantee project success, and in all cases, the user will be billed for cost and labor for UTGC effort as per our fee structure. All samples are assumed to be safe and no greater than BSL1, unless otherwise agreed upon in advance. Contact us at genomicscore@utk.edu.
DNA/RNA extraction
Nucleic acid is extracted and purified
Library Prep
Prepare sample for sequencing by adding appropriate adapters
Sequencing
Read the sequence of the sample and create raw data
Analysis
3 Steps: Primary and secondary analysis, and interpretation
DNA Extraction
Some DNA extractions are suitable for automation and some are better suited for manual extraction. Automated extractions are performed on a KingFisher Flex using Zymo Research kits and manual extractions are performed using Qiagen extraction kits. Please contact Core Facility staff to discuss which method is best for your sample type.
| Cost/Pricing Details | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| Manual DNA extraction, each | $20.00 | $25.00 | $36.00 |
| Automated DNA extraction, 1-48 samples, each | $14.00 | $18.00 | $25.00 |
| Automated DNA extraction, 49-96 samples, each | $9.00 | $13.00 | $20.00 |
Genome and Metagenome Library Preparation
Genome and metagenome libraries are prepared using Illumina’s DNA Prep kit (formerly Nextera DNA Flex Library Prep). The starting material needed is 100-500 ng of DNA in less than 30 uL volume, diluted in Tris-HCl, pH 8. The DNA should have an absorbance of 260/280 in the 1.8-2 range and 260-230 in the 2-2.2 range. Contact Core Facility staff if your template DNA does not meet these specifications.
The costs below are for library construction of extracted, quantified DNA. Next you will need to select the appropriate MiSeq or NovaSeq run.
| Cost/Pricing Details | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| Genome and metagenome preps, 1-11 samples, each | $81.00 | $100.00 | $120.00 |
| Genome and metagenome preps, 12-48 samples, each | $70.00 | $88.00 | $106.00 |
| Genome and metagenome preps, >48 samples, each | $55.00 | $70.00 | $85.00 |
Metabarcoding of Amplicon Products
Amplicon libraries are prepared with Illumina’s two-step PCR Nextera XT amplicon protocol. Users can prepare their own PCR products or Core staff can fully prepare the amplicon libraries with common primers for 16S, ITS, and COI. Please contact Core staff prior to preparing your own library to confirm compatibility with Illumina sequencing platforms, as specific adapters are required. These are the costs for library construction of extracted, quantified DNA. For full library prep, DNA should be diluted to 10 ng/ul, with 10 ul provided. Next you will need to select the appropriate MiSeq or NovaSeq run.
| Cost/Pricing Details | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| Full Amplicon library prep, up to 24 samples | $1,100.00 | $1,375.00 | $1,650.00 |
| Full Amplicon library prep, 25-48 samples | $1,400.00 | $1,750.00 | $2,100.00 |
| Full Amplicon library prep, 49-96 samples | $1,850.00 | $2,300.00 | $2,775.00 |
| Amplicon library prep of post-PCR product, up to 24 samples | $850.00 | $1,065.00 | $1,275.00 |
| Amplicon library prep of post-PCR product, 25-48 samples | $1,100.00 | $1,375.00 | $1,675.00 |
| Amplicon library prep of post-PCR product, 49-96 samples | $1,475.00 | $1,875.00 | $2,215.00 |
RNASeq
RNA libraries are prepared using Zymo Research’s Zymo-Seq RiboFree Total RNA Library Kit, which includes an rRNA depletion step. The minimum input amount of RNA is 100 ng, but the recommended input is 500 ng. RNA should have A260/A280 and A260/A230 ratios >1.8 and be DNA-free. Please check that this kit is compatible with your needs.
The costs below are for library construction of extracted, quantified and quality confirmed RNA. Next you will need to select the appropriate MiSeq or NovaSeq run.
| Cost/Pricing Details | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| RNA prep of up to 12 samples, each | $110.00 | $140.00 | $170.00 |
| RNA prep of 13-48 samples, each | $100.00 | $125.00 | $150.00 |
| RNA prep of >48 samples, each | $86.00 | $105.00 | $125.00 |
Illumina MiSeq Sequencing
The Illumina MiSeq is a benchtop sequencer specialized for lower-output runs on single-lane flow cells, offering Illumina’s longest read length of up to 300 nucleotides, paired-end. The MiSeq is ideal for amplicon sequencing, such as for bacterial 16S rRNA, and small genomes. It can also be used to test larger runs prior to running them on the larger Illumina NovaSeq.
The appropriate Illumina MiSeq flow cell for your project is determined by the depth of coverage needed, as well as the desired read length. Contact Core facility staff for advice on which flow cell is right for your project. The costs below are for a PREPARED library. Costs for library prep done at the Core facility listed on this website are in addition to the cost of the flow cell.
| MiSeq Service | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| v2, 50 cycle flow cell, 2 x 25 nt Sequencing, ~12-14 million read pairs | $1,000.00 | $1,150.00 | $1,350.00 |
| v2, 500 cycle flow cell, 2 x 250 nt Sequencing, ~12-14 million read pairs | $1,600.00 | $1,750.00 | $1,950.00 |
| v3, 150 cycle flow cell, 2 x 75 nt Sequencing, ~22-25 million read pairs | $1,285.00 | $1,435.00 | $1,635.00 |
| v3, 600 cycle flow cell, 2 x 300 nt Sequencing, ~22-25 million read pairs | $1,975.00 | $2,125.00 | $2,325.00 |
| micro cycle flow cell, 2 x 150 nt Sequencing, ~4 million read pairs | $700.00 | $850.00 | $1,050.00 |
| MiSeq run only | $250.00 | $400.00 | $600.00 |
Illumina NovaSeq Sequencing
The NovaSeq 6000 is the latest large-scale sequencer from Illumina, generating unprecedented output in less than two days using patterned flow cell technology, with scalable through-puts to accommodate most projects. Careful consideration of sample numbers and flow cell output is crucial for effective use of this platform. The high yields of the NovaSeq flow cells may present difficulties in efficiently filling flow cells for many researchers. As such, we are currently offering the smallest flow cell, the SP, by single lane use. We hope to expand our options in the future. Custom options may be available upon request. Contact core staff for more information.
The appropriate Illumina NovaSeq flow cell for your project is determined by the depth of coverage needed, as well as the desired read length. Contact Core facility staff for advice on which flow cell is right for your project. The costs below are for a PREPARED library. Costs for library prep done at the Core facility listed on this website are in addition to the cost of the flow cell.
For assistance calculating coverage necessary for your specific project, visit Illumina’s Coverage Calculator.
| NovaSeq Service | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| SP LANE, 2 x 100 nt Sequencing, ~300-400 million read pairs | $2,025.00 | $2,175.00 | $2,375.00 |
| SP LANE, 2 x 150 nt Sequencing, ~300-400 million read pairs | $2,160.00 | $2,310.00 | $2,510.00 |
| SP LANE, 2 x 250 nt Sequencing, ~300-400 million read pairs | $2,800.00 | $2,950.00 | $3,150.00 |
| S1 FLOW CELL, 2 x 150 nt Sequencing, ~1.3-1.6 billion read pairs | $6,000.00 | $6,150.00 | $6,350.00 |
| S2 FLOW CELL, 2 x 150 nt Sequencing, ~3.3-4.1 billion read pairs | $10,635.00 | $10,785.00 | $10,985.00 |
| S4 FLOW CELL, 2 x 150 nt Sequencing, ~8-10 billion read pairs | $15,750.00 | $15,900.00 | $16,100.00 |
| NovaSeq run only | $400.00 | $550.00 | $750.00 |
Applications for Illumina MiSeq and NovaSeq high-throughput sequencing include:
Users may build their own libraries or Genomics Core staff can assist with library preparation. Students are encouraged to participate in lab work associated with their individual projects.
The Oxford MinION sequencer enables direct, real-time analysis of short to ultra-long (>4 Mb) fragments of DNA/RNA in an easy-to-use, low-cost format.
Oxford Nanopore MinION Applications
| Cost/Pricing Details | Internal University of Tennessee User | External academic user | External non-academic user |
|---|---|---|---|
| Library Prep with Oxford Nanopore Rapid Sequencing Kit, per sample | $225.00 | $335.00 | $450.00 |
| Library Prep with Oxford Nanopore Genomic DNA Litigation Kit, per sample | $285.00 | $350.00 | $475.00 |
| Oxford Nanopore MinION run* | $96.00 | $145.00 | $200.00 |
| Oxford Nanopore wash between runs run* | $60.00 | $88.00 | $100.00 |
*Flow cells for the MinION have the potential to be reused but have carryover between runs. For this reason, flow cells must be purchased by each individual user who have the option to reuse their own flow cells. Contact Core Facility staff for more information and flow cell costs.
For further information on applications, project design, and pricing, please contact Veronica Brown at GenomicsCore@utk.edu or 865-974-8080. Please contact Core staff prior to beginning a project to ensure compatibility with the Illumina MiSeq.
For assistance with Illumina data processing:
The CEB Bioinformatics Resource Center (BRC) provides access to computational resources and expert advice for assembling and analyzing Illumina data. The BRC is located in SERF 736.
For further information on applications, project design, and pricing,
please contact Veronica Brown at GenomicsCore@utk.edu or
865-974-8080.
