Development of a Multiplex DNA Typing Method
Location
CSU 253/4/5
Start Date
5-4-2010 10:00 AM
End Date
5-4-2010 12:00 PM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
James Rife
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Short Tandem Repeats (STRs) are locations on DNA where short nucleotide sequences are repeated tandemly. By measuring the number of repeats at specific loci, we can differentiate between individuals. The FBI uses 13 different STR loci on the human genome to link individuals to DNA samples. The objective of this project was to develop a multiplex Polymerase Chain Reaction (PCR) method that could use the LI-COR DNA Analyzer to simulate DNA typing commonly used in forensic labs. If successful, the method could be adapted to teaching labs. Since it was prohibitive to analyze all 13 CODIS STR sites, we focused on four STR loci used by the FBI that would give non-overlapping PCR products. The sites chosen were D13S317, D351385, TPOX, and CSF1PO as well as a sex-determining site, Amelogenin. In an initial analysis, four subjects supplied blood samples. Three of the subjects were siblings. We used a Purelink Genomic DNA Mini Kit from Invitrogen to isolate the DNA and Platinum Taq DNA Polymerase High Fidelity to amplify the DNA. The products of the PCR were analyzed on a Criterion 15% TBE gel. Since this initial analysis demonstrated that the PCR method worked, the samples were further amplified and compared on LI-COR DNA Analyzer to obtain better resolution of the PCR products. The study was expanded to investigate how kinship can be revealed with this method.
Development of a Multiplex DNA Typing Method
CSU 253/4/5
Short Tandem Repeats (STRs) are locations on DNA where short nucleotide sequences are repeated tandemly. By measuring the number of repeats at specific loci, we can differentiate between individuals. The FBI uses 13 different STR loci on the human genome to link individuals to DNA samples. The objective of this project was to develop a multiplex Polymerase Chain Reaction (PCR) method that could use the LI-COR DNA Analyzer to simulate DNA typing commonly used in forensic labs. If successful, the method could be adapted to teaching labs. Since it was prohibitive to analyze all 13 CODIS STR sites, we focused on four STR loci used by the FBI that would give non-overlapping PCR products. The sites chosen were D13S317, D351385, TPOX, and CSF1PO as well as a sex-determining site, Amelogenin. In an initial analysis, four subjects supplied blood samples. Three of the subjects were siblings. We used a Purelink Genomic DNA Mini Kit from Invitrogen to isolate the DNA and Platinum Taq DNA Polymerase High Fidelity to amplify the DNA. The products of the PCR were analyzed on a Criterion 15% TBE gel. Since this initial analysis demonstrated that the PCR method worked, the samples were further amplified and compared on LI-COR DNA Analyzer to obtain better resolution of the PCR products. The study was expanded to investigate how kinship can be revealed with this method.
Recommended Citation
Ranginwala, Mohammad A.. "Development of a Multiplex DNA Typing Method." Undergraduate Research Symposium, Mankato, MN, April 5, 2010.
https://cornerstone.lib.mnsu.edu/urs/2010/poster-session-A/26
