Abstract
Although the dot-blot-SNP technique is a laborsaving, cost-effective method for SNP genotyping of a large number of plants, the synthesis of 5′-digoxigenin (DIG)-labeled oligonucleotides for use as probes is still costly. We developed two probe-labeling methods for this technique, one being digoxigenin labeling of oligonucleotides by PCR (PCR-DIG labeling) and the other being hybridization using a bridge probe and a 5′-DIG-labeled oligonucleotide (bridge hybridization). Bridge hybridization detected allele-specific signals under hybridization conditions similar to those for the 5′-DIG-labeled oligonucleotides and biotin-labeled oligonucleotides, while signals were detected only under a lower stringency condition by PCR-DIG labeling. As a method for genotyping using many markers at one time, two methods, i. e., PCR using mixed primer pairs and hybridization using mixed probes, were examined with successful results. Eighty-five SNP markers designed for genotyping of rice cultivars detected allele-specific signals, the genotyping results corresponding to the previously reported ones.
Original language | English |
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Pages (from-to) | 179-185 |
Number of pages | 7 |
Journal | Molecular Breeding |
Volume | 25 |
Issue number | 1 |
DOIs | |
Publication status | Published - 2010 Dec 1 |
Keywords
- Cultivar identification
- Dot-blotting
- Large-scale genotyping
- SNP analysis
ASJC Scopus subject areas
- Biotechnology
- Molecular Biology
- Agronomy and Crop Science
- Genetics
- Plant Science