SNP Genotyping by Heteroduplex Analysis View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2015

AUTHORS

Norma Paniego , Corina Fusari , Verónica Lia , Andrea Puebla

ABSTRACT

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. More... »

PAGES

141-50

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4939-1966-6_10

DOI

http://dx.doi.org/10.1007/978-1-4939-1966-6_10

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1030851039

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25373754


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