1. PCR amplification of SNP loci from crude DNA for large-scale genotyping of oomycetes
- Author
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Yuxin Zhou, Rebecca Lyon, Kurt Lamour, and Jian Hu
- Subjects
0106 biological sciences ,0301 basic medicine ,Genotype ,Physiology ,Single-nucleotide polymorphism ,Biology ,Molecular Inversion Probe ,Polymerase Chain Reaction ,Polymorphism, Single Nucleotide ,01 natural sciences ,Genome ,law.invention ,03 medical and health sciences ,Nucleic acid thermodynamics ,law ,Genetics ,Molecular Biology ,Genotyping ,Ecology, Evolution, Behavior and Systematics ,Polymerase chain reaction ,Plant Diseases ,Oomycete ,DNA ,Cell Biology ,General Medicine ,biology.organism_classification ,SNP genotyping ,030104 developmental biology ,Oomycetes ,010606 plant biology & botany - Abstract
Similar to other eukaryotes, single nucleotide polymorphism (SNP) markers are abundant in many oomycete plant pathogen genomes. High resolution DNA melting analysis (HR-DMA) is a cost-effective method for SNP genotyping, but like many SNP marker technologies, is limited by the amount and quality of template DNA. We describe PCR preamplification of Phytophthora and Peronospora SNP loci from crude DNA extracted from a small amount of mycelium and/or infected plant tissue to produce sufficient template to genotype at least 10 000 SNPs. The approach is fast, inexpensive, requires minimal biological material and should be useful for many organisms in a variety of contexts.
- Published
- 2014
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