High-resolution SNP mapping by denaturing HPLC |
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| Authors: | Nairz Knud Stocker Hugo Schindelholz Benno Hafen Ernst |
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| Affiliation: | Zoologisches Institut der Universit?t Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland. |
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| Abstract: | With the availability of complete genome sequences, new rapid and reliable strategies for positional cloning become possible. Single-nucleotide polymorphisms (SNPs) permit the mapping of mutations at a resolution not amenable to classical genetics. Here we describe a SNP mapping procedure that relies on resolving polymorphisms by denaturing HPLC without the necessity of determining the nature of the SNPs. With the example of mapping mutations to the Drosophila nicastrin locus, we discuss the benefits of this method, evaluate the frequency of closely linked and potentially misleading second site mutations, and demonstrate the use of denaturing high-performance liquid chromatography to identify mutations in the candidate genes and to fine-map chromosomal breakpoints. Furthermore, we show that recombination events are not uniformly dispersed over the investigated region but rather occur at hot spots. |
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