A robust method for detecting single-nucleotide changes as polymorphic markers by PCR.

S D Michaels; R M Amasino

doi:10.1046/j.1365-313x.1998.00123.x

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PubMed · 9628032

A robust method for detecting single-nucleotide changes as polymorphic markers by PCR.

S D Michaels·R M Amasino

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Abstract

Numerous techniques in plant molecular genetic analysis, such as mapping and positional cloning techniques, rely on the availability of molecular markers that can differentiate between alleles at a particular locus. PCR-based cleaved amplified polymorphic sequences (CAPS) markers have been widely used as a means of rapidly and reliably detecting a single-base change that creates a unique restriction site in one of a pair of alleles. However, the majority of single-nucleotide changes do not create such sites and thus cannot be used to create CAPS markers. In this paper, a modification of the CAPS technique that allows detection of most single-nucleotide changes by utilizing mismatched PCR primers is described. The mismatches in the PCR primers, in combination with the single-nucleotide change, create a unique restriction site in one of the alleles.

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Base Sequence DNA Primers Deoxyribonucleases, Type II Site-Specific Genetic Markers Point Mutation Polymerase Chain Reaction Polymorphism, Genetic Restriction Mapping Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

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S D Michaels, R M Amasino. 1998. A robust method for detecting single-nucleotide changes as polymorphic markers by PCR.. https://doi.org/10.1046/j.1365-313x.1998.00123.x

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