SNP genotyping by heteroduplex analysis
Heteroduplex-based genotyping methods have proven to be technologically effective and economically efficient for low- to medium-range throughput single-nucleotide polymorphism (SNP) determination. In this chapter we describe two protocols that were successfully applied for SNP detection and haplotype analysis of candidate genes in association studies. The protocols involve (1) enzymatic mismatch cleavage with endonuclease CEL1 from celery, associated with fragment separation using capillary electrophoresis (CEL1 cleavage), and (2) differential retention of the homo/heteroduplex DNA molecules under partial denaturing conditions on ion pair reversed-phase liquid chromatography (dHPLC). Both methods are complementary since dHPLC is more versatile than CEL1 cleavage for identifying multiple SNP per target region, and the latter is easily optimized for sequences with fewer SNPs or small insertion/deletion polymorphisms. Besides, CEL1 cleavage is a powerful method to localize the position of the mutation when fragment resolution is done using capillary electrophoresis.
| Main Authors: | , , , |
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| Format: | info:ar-repo/semantics/artículo biblioteca |
| Language: | eng |
| Published: |
Springer
2015-10
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| Subjects: | Single Nucleotide Polymorphism, Electrophoresis, Genotypes, Polimorfismo de un Solo Nucleótido, Electroforesis, Genotipos, Candidate Genes, Heteroduplex Analysis, Genes Candidatos, Análisis heterodúplex, |
| Online Access: | http://hdl.handle.net/20.500.12123/8118 https://link.springer.com/protocol/10.1007/978-1-4939-1966-6_10 https://doi.org/10.1007/978-1-4939-1966-6_10 |
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