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Random amplified polymorphic DNA (RAPD) sequences as markers for canine genetic studies
1James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
The Journal of Heredity
|February 13, 1999
Summary
Random amplified polymorphic DNA (RAPD) markers show promise for canine genetic studies. However, using specific primer pairs to amplify RAPD sequences can lead to a loss of polymorphism in some cases.
Area of Science:
- Molecular genetics
- Canine genetics
Background:
- Random amplified polymorphic DNA (RAPD) markers are utilized in genetic studies.
- Reproducibility issues with RAPD-PCR reactions have been reported.
Purpose of the Study:
- To apply the RAPD-PCR assay to canine genetic studies.
- To investigate the amplification of RAPD sequences using specific primer pairs.
Main Methods:
- Tested 240 decanucleotide primers for PCR amplification.
- Cloned and sequenced five polymorphic RAPD fragments.
- Synthesized specific 24mer primer pairs for characterized fragments.
- Analyzed amplification products using restriction digests and Southern analysis.
Main Results:
- 34.6% of primers yielded polymorphic fragments in a Labrador retriever pedigree.
- Two specific primer pairs amplified exclusively positive RAPD fragment sequences.
- Three specific primer pairs amplified sequences from all DNA samples, losing polymorphism.
- A new MseI restriction polymorphism was identified in one amplification product.
- Cloned sequences did not contain long open reading frames and showed species-specific hybridization.
Conclusions:
- RAPD sequences can serve as molecular markers for canine genetic studies.
- Employing specific primer pairs for RAPD sequences may result in loss of polymorphism for some markers.