Take a look at the DSC's Guide to NGS Project Planning. Whether you're new to NGS or a returning user, this guide will help you plan your project effectively, from sample collection to data output.
Step One: Get in touch and get a quote
We always recommend taking advantage of our free consultation services before starting your project. Contact us, and we’ll talk to you about the best preparation and sequencing methods for your project, and how to prepare samples to maximize your chances of success.
Your next step will need to be getting a quote. For groups outside of the University of Toronto, we require institutional purchase orders (POs) for payment, and your procurement department will need a quote from us to issue one. Download the Sample Information Sheet, fill out at least the top section of the Project Info & Checklist tab, and email it to firstname.lastname@example.org with the subject line Quote Request. Or, send that Quote Request email with the following information:
Lead PI’s full name and affiliation (whoever will be funding the project)
Project type: User-prepared libraries, or samples for prep and sequencing
Library preparation type requested
Read length (preferrred and acceptable alternatives)
Number of read clusters/sample
Sample type and species
High (>65%) GC/AT content?
Sequence complexity (low/high)
Check the Project Planning Guide (see Section 6) for help in selecting these parameters.
We know that you’re operating within budgetary constraints; let us know your situation. This can help us select options to maximize the data you get with the money you have.
Quotes last for 3 months after issue. Ask your procurement department to generate a purchase order, which we’ll require before completing your project. After sequencing and data distribution, you’ll receive an official invoice from the University of Toronto. Make sure that the invoice documentation makes it back to your procurement department so payment is completed.
Step Two: Prepare your samples
Make sure you take a look at our sample submission requirements below before extracting or preparing your material.
If you’re submitting user-prepared libraries, we can do quality-control and pooling, or take samples pooled by you. Libraries and pools should be in water or 10 mM Tris. Pools should ideally be at least 5 nM, in the following minimum volumes:
- MiSeq: 10 uL
- NextSeq: 20 uL
- NovaSeq SP/S1: 125 uL
- NovaSeq S2: 175 uL
- NovaSeq S4: 335 uL
Contact us if your libraries or pools do not meet these minima; we can often make it work. Please note that we cannot guarantee sequencing performance of customer-prepared libraries, but we’re always happy to consult with you on how to maximize your chances of success.
For library preparation at the DSC, we ask that your material be:
Sufficient – Check Table I in the Project Planning Guide for minimum amounts of submitted material for each of our services. Note that NanoDrop and similar instruments are not considered to be reliable for quantification; use a fluorometric method like Qubit or Quant-iT. Contact us if you need us to run this quantification for you. Note that the listed quantities are minimums – for best results, submit at least double these amounts.
Clean – Contaminants from extractions can interfere with library preparation; that’s why we require that all material be submitted in molecular grade water or 10 mM Tris pH 8.0. We also require that you test for chemical contamination on a NanoDrop and state the 260/280 and 260/230 ratios in your submission file.
Intact – Degraded material can also cause problems for library preparation and negatively impact data quality. That’s why we require RIN scores over 8 for most RNA samples, and need to see gel images for DNA. We also offer BioAnalyzer service if you’re unable to determine RIN scores yourself.
We’re flexible – if your samples don’t meet some of our requirements, or if you’re unable to submit NanoDrop values or gel images, we’re happy to proceed with the preparation. However, samples with low or unknown starting quality are considered “on risk”, and the customer will still be financially liable if preparation fails.
Step Three: Submit!
We accept samples in clearly-labelled strip tubes or 96-well plates. Pooled libraries may be submitted in 1.5-mL tubes. Make sure you’ve completely filled out the Project Info & Checklist and Sample Information tabs of the Sample Information Sheet, taking care to match what’s written in the form with what’s written on the tubes. The completed sheet must be emailed to us before we can complete your project. For groups outside of the University of Toronto, we'll also need to receive your purchase order - in the interest of time, we can start library prep without this PO, but will not complete sequencing until we receive it.
If you’re local, you can drop off samples directly with us at the DSC. If you’re further away, ship with sufficient dry ice to last through transit. If shipping internationally, always ship on a Monday to leave time for Customs clearance.
University of Toronto
Donnelly Sequencing Centre
Attention: Tanja Durbic
160 College Street, Room 643
Toronto, Ontario, M5S 3E1
416.978.8579 | email@example.com
Submission of samples confirms acceptance of the terms and conditions outlined in the Sample Information Sheet. We look forward to working with you!