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