Haplotyping using a combination of polymerase chain reaction–single-strand conformational polymorphism analysis and haplotype-specific PCR amplification
Abstract
A single nucleotide polymorphism (SNP) may have an impact on phenotype, but it may also be influenced by multiple SNPs within a gene; hence, the haplotype or phase of multiple SNPs needs to be known. Various methods for haplotyping SNPs have been proposed, but a simple and cost-effective method is currently unavailable. Here we describe a haplotyping approach using two simple techniques: polymerase chain reaction–single-strand conformational polymorphism (PCR–SSCP) and haplotype-specific PCR. In this approach, individual regions of a gene are analyzed by PCR–SSCP to identify variation that defines sub-haplotypes, and then extended haplotypes are assembled from the sub-haplotypes either directly or with the additional use of haplotype-specific PCR amplification. We demonstrate the utility of this approach by haplotyping ovine FABP4 across two variable regions that contain seven SNPs and one indel. The simplicity of this approach makes it suitable for large-scale studies and/or diagnostic screening.... [Show full abstract]
Keywords
haplotyping; SNP haplotype; PCR–SSCP; haplotype-specific PCR amplification; PCR-SSCP; Biochemistry & Molecular Biology; Animals; Sheep; Humans; DNA Fingerprinting; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; Base Sequence; Haplotypes; Polymorphism, Single-Stranded Conformational; Molecular Sequence Data; Fatty Acid-Binding ProteinsFields of Research
060101 Analytical BiochemistryDate
2014-12-01Type
Journal ArticleCollections
- Metadata-only (no full-text) [4903]
© 2014 Elsevier Inc. All rights reserved.
Citation
Zhou et al. (2014). Haplotyping using a combination of polymerase chain reaction–single-strand conformational polymorphism analysis and haplotype-specific PCR amplification. Analytical Biochemistry, 466, 59-64. doi:10.1016/j.ab.2014.08.021Related items
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