SUBMITTING SAMPLES

Samples can be submitted for genotyping using the following process and guidelines:

Process

1

Enter your order request online using dnaLIMS

Choose the correct filter set based on the appropriate size standard and include the requested size standard in the Comments section.

Please contact us to ensure we have the required size standard in stock

2

Submit samples with a printed copy of your request form

  • A minimum of 5µl labeled PCR products are submitted (8-strip tubes or 96-well plate - see the orientation grid)
  • Please contact us to discuss processing requirements for 384-well plates

3

Drop off in-person, by campus mail, or by courier to:

Sequencing and Bioinformatics Consortium
University of British Columbia
Room 3124 - PharmSci Building
2405 Wesbrook Mall
Vancouver, BC V6T 1Z3

Samples can be dropped off at any time in the refrigerated dropbox on the main floor of the Pharmaceutical Sciences Building. Contact us for more information.  


Size Standards

An appropriate size standard is chosen based on the expected fragment sizes for the project, and it is imperative that the same size standard be used for the duration of that project. Based on the Local Southern Size-Calling Method, there must be two size standard peaks smaller than the shortest fragment, and two size standard peaks larger than the longest fragment.

Local Southern Size-calling Method

  • A curve is created by using three standard points (two points below and one point above the fragment), then a fragment size is determined.
  • Another curve is created by looking at an additional set of three points (one point below and two points above the fragment), then another value is assigned
  • The two size values are averaged to determine the unknown fragment l

The fluorophore label on the size standard determines the level of multiplexing available. The ROX™ size standards can accomodate sample fragments labeled with up to three different fluorophores, whereas the LIZ™ size standards permit multiplexing with four different fluorophores.

SBC maintains stocks of the following size standards:

4-COLOUR FLUORESCENCE – each DNA fragment labeled with ROX™ Fluorophore

5-COLOUR FLUORESCENCE – each DNA fragment labeled with LIZ® Fluorophore


Filter Sets
 

 

Genotyping-Filter-Sets.png

Orientation Grid

 

 

96-well-plate.PNG

 

96-well-plate-orientation.png

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