Donnelly Centre for Cellular and Biomolecular Research

Getting Started

Download a checklist of your next steps here.

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 dseqcentre@utoronto.ca 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)

  • Number of samples for library preparation

  • Library preparation type requested

  • Number of samples for sequencing

  • Read length

  • Number of reads/sample

  • Desired sequencing instrument and acceptable alternatives

  • Sample type and species

  • High (>65%) GC/AT content?

  • Sequence complexity (low/high)

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 samples that you prepared for sequencing, libraries must be cleaned to remove adapter- and primer-dimers, and must be suspended in molecular grade water or 10 mM Tris pH 8.0 – never in TE or similar buffers. Contact us about your sequencing needs and we'll let you know how much material to submit. 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 – See the chart below for our offered services and the minimum mass of sample that we’ll need. 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.

 

Method Preparation kit Min. mass (ng) Min. conc. (ng/uL)
RNA: Stranded mRNA-Seq NEBNext Ultra II Directional
NEBNext DSC Long-Read Protocol
TruSeq Stranded mRNA
TruSeq DSC Long-Read Protocol
10
20
100
200
0.2
0.4
2
4
RNA: Non-stranded mRNA-Seq SMART-Seq v4 Ultra-Low + Nextera XT
SMART-Seq v4 Ultra-Low + Nextera XT
0.01
1 cell
0.002
0.2 cells/uL
RNA: 3' RNA-Seq SureCell WTA 3'
SMART-Seq v4 3' DE + Nextera XT
SMART-Seq v4 3' DE + Nextera XT
11250 cells
0.01
1 cell
2500 cells/uL
0.002
0.2 cells/uL
RNA: Stranded Ribo-depletion RNA-Seq NEBNext Ultra II Directional
NEBNext Ultra II Directional for FFPE
NEBNext rRNA-Dep + SMARTer Stranded
NEBNext rRNA-Dep + SMARTer Univ Low
TruSeq Stranded Total Ribo-Zero
5
10
4
10
100
0.42
0.84
0.34
0.84
10
RNA: Small RNA-Seq NEB Small RNA
SMARTer smRNA-Seq
100
1
17
0.17
RNA: Targeted RNA-Seq SMARTer Target RNA Capture + Nextera
TruSeq RNA Exome (stranded)
10
10*
2
1.2
DNA: Shotgun sequencing NEBNext Ultra II DNA
Nextera DNA Flex
Nextera DNA XT
NEB FFPE + NEBNext Ultra II DNA
0.5
1
1
5
0.01
0.034
0.2
0.11
DNA: Amplicon sequencing and 16S rRNA Amplicon PCR sequencing
16S rRNA metagenomics sequencing
1
12.5
0.1
2.5
DNA: Mate-pair sequencing Nextera Mate Pair 4000 13
DNA: Whole-exome sequencing TruSeq DNA Exome
Nextera DNA Exome
Nextera Flex for Enrichment
SureSelect XT2 Exome
100
50
10
1000
2
5
0.34
20
DNA: ChIP-Seq NEBNext Ultra II DNA for ChIP
TruSeq ChIP-Seq
DNA SMART ChIP-Seq
0.5
5
0.1
0.01
0.1
0.005
DNA: Methylation EpiMark Methylated DNA + Ultra II DNA
TruSeq Methyl Capture EPIC
TruSeq DNA Methylation
EpiXplore Meth-Seq
5
500
50
50
0.1
10
5.6
1
DNA: CRISPR Custom methods (single and dual guides) 10 2

*Degraded or FFPE samples will require higher inputs.

Step Three: Submit!

We accept samples in 1.5 mL tubes, strip tubes, or 96-well plates. 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 | dseqcentre@utoronto.ca

Submission of samples confirms acceptance of the terms and conditions outlined in the Sample Information Sheet. We look forward to working with you!