A robust method for detecting single‐nucleotide changes as polymorphic markers by PCR |
| |
Authors: | Scott D Michaels & Richard M Amasino |
| |
Institution: | Department of Biochemistry, University of Wisconsin‐Madison, 420 Henry Mall, Madison, WI 53706, USA |
| |
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. |
| |
Keywords: | |
|
|