Services
Illumina sequencing
We offer Illumina library preparation services using commercially available kits. We have two Illumina sequencing instruments: NextSeq 2000 and MiniSeq. Our sequencing output has a range of 8 million to 1.8 billion read/read pairs, or 2.4 to 540 gigabases. This is the equivalent of 0.6 to 110 human genome equivalents.
| Instrument | Kit type | Cycles | Advertised Gb | Cost/Gb | Cost/1M reads/read pairs | Human Genome equivalents |
|---|---|---|---|---|---|---|
| NextSeq 2000 | P1 | 100 | 10 | $68.30 | $6.84 | 3.3 |
| NextSeq 2000 | P1 | 300 | 30 | $31.67 | $9.50 | 10 |
| NextSeq 2000 | P1 | 600 | 60 | $24.07 | $14.44 | 20 |
| NextSeq 2000 | P2 | 100 | 40 | $25.94 | $2.59 | 13.3 |
| NextSeq 2000 | P2 | 200 | 80 | $24.38 | $4.88 | 26.7 |
| NextSeq 2000 | P2 | 300 | 120 | $21.55 | $6.46 | 40 |
| NextSeq 2000 | P2 | 600 | 240 | $11.73 | $7.04 | 80 |
| NextSeq 2000 | P3 | 100 | 120 | $18.46 | $1.85 | 40 |
| NextSeq 2000 | P3 | 200 | 240 | $12.78 | $2.56 | 80 |
| NextSeq 2000 | P3 | 300 | 360 | $11.36 | $3.41 | 120 |
| NextSeq 2000 | P4 | 50 | 90 | $19.00 | $0.95 | 30 |
| NextSeq 2000 | P4 | 100 | 180 | $14.25 | $1.43 | 60 |
| NextSeq 2000 | P4 | 200 | 360 | $10.08 | $2.02 | 120 |
| NextSeq 2000 | P4 | 300 | 540 | $8.55 | $2.57 | 180 |
| Instrument | Kit type | Cycles | Sequencing kit cost (w/ shipping) | Advertised reads/read pairs (millions) | Advertised Gb | Cost/Gb | Cost/1M reads/read pairs | Human Genome equivalents |
|---|---|---|---|---|---|---|---|---|
| MiniSeq | Rapid high output | 100 | $1,083.53 | 20 | 2 | $498.75 | $49.88 | 0.7 |
| MiniSeq | High output | 75 | $949.60 | 25 | 1.88 | $464.89 | $34.96 | 0.6 |
| MiniSeq | High output | 150 | $1,109.71 | 25 | 3.75 | $272.33 | $40.85 | 1.3 |
| MiniSeq | High output | 300 | $1,780.38 | 25 | 7.5 | $218.50 | $65.55 | 2.5 |
| MiniSeq | Mid output | 300 | $634.41 | 8 | 2.4 | $243.44 | $73.03 | 0.8 |
* Yields are as advertised by Illumina. Results will vary.
Contact the ÁùºÏ±¦µä Genomics Center to begin an Illumina sequencing project.
Sanger sequencing
The ÁùºÏ±¦µä Genomics Center operates a SeqStudio Genetic Analyzer for Sanger sequencing. All of our runs include an internal plasmid control to aid in quality control.
Sanger results (.AB1 files) will be returned to clients in 24-48 hours.
- Follow the guidelines for the optimal amount of template and primer for Sanger sequencing..
- We request a minimum volume of 5uL; if your sample is less than 5uL please add water to bring the submitted sample up to 5uL. The maximum volume we accept is 25uL, so if your template DNA is too dilute please concentrate it.
- Submitted samples should be in water. EDTA is common in elution buffers and will inhibit the Sanger reaction.
- Do NOT use DEPC-treated water for any dilutions involved in Sanger sample preparation.
- For PCR amplicons:
- Ensure only one PCR product is present (visualize with a gel).
- PCR reactions must be cleaned up prior to sequencing using routine cleanup kits. We offer PCR cleanup services.
- Use a dsDNA-specific quantification method (e.g. Qubit, picogreen) to quantify your templates. We offer quantification services.
- How to submit your samples:
- Using 8-well strip tubes:
- Label each tube with the sample number (1, 2, 3, …).
- Write the order number on the side of the first tube of the strip.
- Using 96-well plates:
- Submit a maximum of 96 samples per order if you are using plates.
- Order samples down columns. Sample 1 is A1, sample 2 is B1, etc.
- Write the order number on the plate.
- Email the Sanger submission form to the ÁùºÏ±¦µä Genomics Center prior to dropping off samples.
- Samples may be physically submitted to ARF 408, or mailed to the ÁùºÏ±¦µä Genomics Center.
- There is a Sanger drop-off basket in the hallway if the laboratory is closed. Samples will be stable if left overnight.
- Using 8-well strip tubes:
Fragment analysis/genotyping/DNA fingerprinting
Fragment analysis can be performed using microsatellite or RFLP techniques. Samples are run on the SeqStudio Genetic Analyzer, which is equipped with filters compatible with 6-FAM, VIC, NED, PET and LIZ (for size standards).
We can run plates provided as-is, or prepare your samples by diluting (if necessary) and adding size standard (LIZ 600).
We will determine the best dilution for your samples if necessary for a nominal fee.
- Ensure that your sample prep process (PCR) is working optimally and robustly. We recommend being able to dilute PCRs by at least 100-fold for optimal capillary runs.
- How to submit your samples:
- Using 8-well strip tubes:
- Label each tube with the sample number (1, 2, 3,…).
- Write the order number on the side of the first tube of the strip.
- Using 96-well plates:
- Submit a maximum of 96 samples per order if you are using plates.
- Order samples down columns. Sample 1 is A1, sample 2 is B1, etc.
- Write the order number on the plate.
- Using 8-well strip tubes:
PCR cleanup
We offer PCR cleanup for Sanger sequencing services. Your samples will be quantified and an appropriate amount is used in an enzymatic reaction that degrades remaining PCR primers and dephosphorylates remaining dNTPs. This treated sample can then be used directly in Sanger sequencing.
How to submit your samples:
- Using 8-well strip tubes:
- Label each tube with the sample number (1, 2, 3,…).
- Write the order number on the side of the first tube of the strip.
- Using 96-well plates:
- Submit a maximum of 96 samples per order if you are using plates.
- Order samples down columns. Sample 1 is A1, sample 2 is B1, etc.
- Write the order number on the plate.
Quantification services
We use quantification methods specific for dsDNA or RNA.
Up to 48 samples will be quantified with a Qubit fluorometer.
A plate-based assay will be used to quantify 48 or more samples. Standards are measured in triplicate and you can request multiple measurements of your samples.
We must be able to pipet a minimum of 2 ul of your samples; provide 3 ul.
How to submit your samples:
- Using 8-well strip tubes:
- Label each tube with the sample number (1, 2, 3,…).
- Write the order number on the side of the first tube of the strip.
- Using 96-well plates:
- Submit a maximum of 96 samples per order if you are using plates.
- Order samples down columns. Sample 1 is A1, sample 2 is B1, etc.
- Write the order number on the plate.
You will receive a spreadsheet listing the concentration of your samples. If you requested a plate-based assay for quantification, the data, including standard curve and subsequent calculations, will be included.
Agilent TapeStation 4150 services
The Agilent TapeStation 4150 can be used to determine the size range of DNA or RNA samples. RIN numbers are reported for RNA samples. We are currently offering three types of ScreenTapes as part of our regular services. Please contact us if you are interested in the High Sensitivity versions of the DNA & RNA ScreenTapes, as well as cell-free and genomic (up to 60 kb) DNA ScreenTapes.
TapeStation D1000
Samples should be 0.1 to 50 ng/ul. The dynamic size range is 35 to 1,000 bp.
TapeStation RNA
Samples should be 25 to 500 ng/ul.
Runs require 1-2 uL of sample, but please submit a minimum 3uL aliquot so that we may accurately remove a 1-2 uL aliquot. We can process any number of samples. Please submit samples in strip tubes.
A PDF report with the results will be emailed to you.
QuantStudio 3 services
We have a QuantStudio 3 that can be used with 0.2 ml PCR plates.
Contact the ÁùºÏ±¦µä Genomics Center to discuss use of the QuantStudio 3. In general, you will need to prepare plates yourself and bring them to the genome center to run. You will be able to easily set up and access your qPCR runs via ThermoFisher Connect, a cloud-based application.