A high-throughput protocol for extracting high-purity genomic DNA from plants and animals |
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Authors: | Whitlock R Hipperson H Mannarelli M Burke T |
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Institution: | Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK. |
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Abstract: | DNA extraction techniques that employ the reversible binding of DNA to silica via chaotropic salts can deliver high-quality genomic DNA from plant and animal tissues, while avoiding the use of toxic organic solvents. Existing techniques that use this method are either prohibitively expensive, or are applicable to only a restricted set of taxa. Here we describe a cost-effective DNA extraction technique suitable for a wide range of plant and animal taxa that yields microgram quantities of high-molecular-weight genomic DNA at a throughput of 192 samples per day. Our technique is particularly robust for tissue samples that are insoluble or are rapidly discoloured or oxidized in standard DNA extraction buffers. We demonstrate the quality of DNA extracted using this method by applying the amplified fragment length polymorphism technique to plant species. |
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Keywords: | automation chaotropic salt DNA extraction high‐throughput microtitre‐plate nucleic acid extraction recalcitrant tissue samples silica |
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