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General approach to analysis of polymorphic short tandem repeat loci
C J Sprecher1, C Puers, A M Lins
1Promega Corporation, Madison, WI, USA.
Biotechniques
|February 1, 1996
Summary
This study presents an affordable, non-isotopic DNA identification method using silver-stained short tandem repeat (STR) analysis. This technique is valuable for forensic science, paternity testing, and disease detection.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Short tandem repeat (STR) loci are crucial for DNA identification.
- Traditional non-isotopic detection methods can be costly or complex.
- Developing efficient and artifact-free amplification and detection systems is essential.
Purpose of the Study:
- To develop an inexpensive and general non-isotopic analytical method for DNA identification.
- To select optimal short tandem repeat (STR) loci for reliable analysis.
- To establish allelic ladders for accurate STR profiling.
Main Methods:
- Polymerase chain reaction (PCR) amplification of 22 known polymorphic STR loci.
- Denaturing polyacrylamide gel electrophoresis (PAGE) for fragment separation.
- Silver staining for sensitive and cost-effective detection of DNA fragments.
Main Results:
- Efficient amplification and minimal artifacts were observed for selected STR loci using silver staining.
- Development of allelic ladders for accurate genotype determination.
- Demonstration of a cost-effective, non-isotopic DNA identification method.
Conclusions:
- The combination of allelic ladders and silver stain detection offers a practical and economical approach to DNA identification.
- This method has broad applications in forensics, paternity testing, cell line identification, and monitoring transplants.
- The approach is adaptable for broader genetic analysis, including disease detection in various species.