- Course Introduction
- 01: Evidence & DNA
- 02: Forensic Biology
- 03: DNA Extraction & Quantitation
- 04: DNA Amplification
- 05: Amplified DNA Product Separation
- 06: STR Data Analysis & Interpretation
- Data Troubleshooting
- Data Interpretation & Allele Calls
The analyst selects one of four possible size calling methods (in the analysis parameters) utilized by GeneScan® :
The Local Southern method for size calling is the most commonly used algorithm in forensic DNA analysis. It determines the sizes of fragments by using the reciprocal relationship between fragment length and mobility. The unknown fragment is surrounded by two known sized fragments above and one below then two below and one above. The results are averaged and the size of the allele is determined.
For a review of size calling methods 2-4 listed above, reference the GeneMapper® ID software version 3.1 User’s Guide.02
Below is a visual representation of the sizing process of the Local Southern method.
In variable-temperature environments, some studies have found that the Global Southern method (rather than the Local Southern method), may provide better sizing precision.03, 04
One important issue concerning the sizing of AmpFℓSTR® GS500 size standards (ROX or LIZ) is the de-labeling of the 250 base pair fragment. At a minimum, the 250 fragment should not be labeled.04
Click here to read more about delabeling in Subject 06, Module 03.
The size calling method is one component of the overall parameter settings that are configured prior to analyzing data.
