A High-Throughput DNA Extraction Protocol for Tropical Molecular Breeding Programs

    Mace, E S and Buhariwalla, H K and Crouch, J H (2003) A High-Throughput DNA Extraction Protocol for Tropical Molecular Breeding Programs. Plant Molecular Biology Reporter, 21 (4). pp. 459-460. ISSN 1572-9818

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    Abstract

    Liquid handling robotics and capillary electrophoresis genetic analyzers now offer high-throughput solutions for 2 of the 4 key steps in PCR-based DNA marker-assisted fingerprinting (DNA extraction, PCR amplification, electrophoresis, data analysis). Thus, DNA extraction remains the most significant bottleneck at the bench for large-scale applications in plant breeding and germplasm characterization. We report on a rapid and low-cost method for relatively high-throughput extraction of high-quality DNA from young and mature leaves of sorghum, pearl millet, chickpea, groundnut, and pigeonpea. The procedure uses a modified CTAB/β-mercaptoethanol method for DNA extraction in a 96-well plate. The quantity and quality of the DNA extracted per sample is adequate for more than 1000 PCR reactions. A relatively high throughput of 96-384 samples per person per day can be achieved, depending on the crop. A major timesaving aspect of the protocol is the absence of a manual sample-grinding step. Finally, the cost is a magnitude lower than commercial plate-based kits, and, as such, is likely to have substantial application in tropical molecular breeding programs.

    Item Type: Article
    Divisions: UNSPECIFIED
    CRP: UNSPECIFIED
    Subjects: Others > Genetics and Genomics
    Others > Agriculture-Farming, Production, Technology, Economics
    Depositing User: Library ICRISAT
    Date Deposited: 28 Dec 2011 06:14
    Last Modified: 28 Dec 2011 06:14
    URI: http://oar.icrisat.org/id/eprint/5093
    Official URL: Http://dx.doi.org/10.1007/BF02772596
    Projects: UNSPECIFIED
    Funders: UNSPECIFIED
    Acknowledgement: UNSPECIFIED
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