Instrumentation
The Donnelly Sequencing Centre is a full service, Illumina CSPro Core sequencing provider.  Our facility utilizes the latest technology including the Illumina HiSeq2500 (upgrade complete November 2012), and MiSeq V2 instruments.  Our quality control services guarantee in-house libraries meet Illumina technical specifications.  Let us help you!  Consultations are free, and help ensure your libraries achieve maximum performance across our platforms.  Please contact Dax Torti (dtorti26@gmail.com,416.978.8579) for further inquiries. 


Library preparation services
We offer complete library preparation for your experimental needs.  Please see required input material recommendations for sample submissions, and suggested quality control procedures listed below.

Genomic DNA Submissions
All libraries should be suspended in molecular grade water, or 10 mM Tris pH 8.0.  We require the following minimum DNA amounts (RNase treated) for each of the listed library preparation methods:

  • Illumina Nextera XT kit sample preparations- 2ng required, recommended  minimum of 100ng
  • Illumina Nextera DNA kit sample preparations – 50 ng required, recommended minimum of 100ng
  • Illumina TruSeq DNA v2 sample preparations- 1ug required, recommended minimum of 2ug
  • Illumina TruSeq Mate Pair v2 sample preparations-  20ug recommended
  • Illumina TruSeq Exome enrichment protocols- 1ug required, recommended minimum 2 ug
  • Illumina ChIP Sequencing preparations-  10ng required, recommended minimum 20 ng
*All libraries subject to internal quality control before library preparation begins

RNA Submissions
All libraries should be suspended in molecular grade water, or 10 mM Tris pH 8.0.  We require the following minimum RNA amounts (DNase treated) for each of the listed library preparation methods:

  • Illumina TruSeq RNA v2 sample preparations- 0.1 ng-4ug total RNA required, recommended minimum 8 ug total RNA
  • Illumina TruSeq stranded mRNA sample preparations- 0.1ng- 1ug total RNA required, recommended minimum of 2 ug total RNA
  • Illumina TruSeq stranded mRNA sample preparations- 0.1ng- 1ug total RNA required, recommended minimum of 2 ug total RNA
  • Illumina TruSeq Small RNA sample preparations-1-10 ug total RNA required, recommended minimum 2 ug total RNA
*All libraries subject to internal quality control before library preparation begins

Prepared Library Submissions
We require a minimum of 40 nmoles of adapter/primer-free library.   We will conduct internal quality control prior to running the library.  We recommend KAPA Library quantification kits (D-Mark) for yield assessment.
 


Quality control

Quality control ensures NGS libraries perform according to illumina standards. The Donnelly Sequencing Centre verifies the integrity of all submitted samples. DNA samples are checked on a gel, and assessed for intact, high molecular weight material.  RNA samples will be subject to Agilent bioanalyzer quality assessment.  Illumina recommends RNA integrity numbers (RIN) greater than 8.  Please follow submission guideline amounts for individual libraries.  We recommend quantitating input material using fluorescent methods for both DNA and RNA (Qubit Quant-it dsDNA HS Assay, and Quant-it RNA assay, Invitrogen).  Nanodrop quantitation is not accurate.  If using this method, please submit recommended minimum material.  Clients will be contacted if samples do not meet specifications. 

Sample Shipment
Sample submission is a simple process.  Fill out our sample submission sheet, including shipment tracking ID, and e-mail the completed form to dtorti26@gmail.com.  All shipments should be on dry ice and addressed as follows:

Dax Torti
Donnelly Sequencing Centre (DSC)
University of Toronto
160 College Street, Room 643
Toronto, Ontario, M5S 3E1
416.978.8579