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Application of DNA Fingerprinting using the D1S80 Locus in Lab Classes
Meike Siebers1, Agatha Walla2, Thea Rütjes3
1Institute of Plant Genetics, Heinrich-Heine-University; meike.siebers@hhu.de.
Journal of Visualized Experiments : Jove
|August 2, 2021
Summary
This study presents a practical DNA fingerprinting method using Variable Number of Tandem Repeat (VNTR) analysis for undergraduate labs. Students learn DNA extraction, PCR, and gel electrophoresis to analyze genetic variation at the D1S80 locus.
Area of Science:
- Biological Sciences
- Genetics
- Forensic Science
Background:
- DNA fingerprinting is crucial in forensics, paternity testing, and evolutionary studies.
- Variable Number of Tandem Repeats (VNTRs) are highly polymorphic genetic markers.
- Implementing hands-on DNA analysis in undergraduate education enhances learning.
Purpose of the Study:
- To describe a reliable and robust method for individual genotyping using VNTR analysis.
- To provide an accessible DNA fingerprinting protocol for undergraduate laboratory classes.
- To educate students on DNA extraction, PCR amplification, and gel electrophoresis techniques.
Main Methods:
- DNA extraction from buccal mucosa epithelial cells.
- Polymerase Chain Reaction (PCR) amplification of the human D1S80 VNTR locus.
- Agarose gel electrophoresis to separate PCR products by size.
- Analysis of fragment sizes and repeat numbers using a DNA size standard and linear regression.
Main Results:
- Successful genotyping of individuals based on D1S80 VNTR locus polymorphisms.
- Demonstration of genetic variation through differences in PCR amplicon sizes.
- Accurate determination of VNTR repeat numbers via gel electrophoresis and regression analysis.
Conclusions:
- The described protocol offers a valuable tool for teaching DNA fingerprinting in undergraduate biology labs.
- Students gain practical skills in molecular biology techniques and data interpretation.
- This method effectively illustrates the application of VNTR analysis in understanding genetic diversity.
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