Low-Cost Methods for Molecular Characterization of Mutant Plants

<p>1             Introduction</p><p>1.1 Background</p><p>1.2 Methods used to isolate genomic DNA from plant tissues</p><p>1.3 Methods for the discovery and characterization of induced and natural nucleotide variation in plant genomes</p><p>References</p><p>2             Health and Safety Considerations</p><p>               2.1 Guidelines</p><p>               2.2 Preparation of a home-made chemical spill kit</p><p>References</p><p>3             Sample Collection and Storage</p><p>3.1 Background</p><p>3.2 Materials</p>3.3 Methods<p><p>       References</p><p>4             Low-Cost DNA Extraction</p><p>               4.1 Materials</p><p>   4.2 Methods</p><p>   4.3 Alternative buffers for DNA extraction</p><p>   References</p><p>5             PCR Amplification for Low-Cost Mutation Discovery</p><p>               5.1 Materials</p><p>               5.2 Methods</p><p>               References</p><p>6             Enzymatic Mismatch Cleavage and Agarose Gel Evaluation of Samples</p>6.1 Materials<p><p>6.2 Methods</p><p>References</p><p>7          Alternative Enzymology for Mismatch Cleavage for TILLING and Ecotilling: Extraction of Enzymes form Common Weedy Plants</p><p>7.1 Materials</p><p>7.2 Methods</p><p>8          Example Data</p><p>8.1 Quality of genomic DNA obtained by silica powder-based DNA extraction method</p><p>8.2 Quality of genomic DNA obtained by silica powder-based DNA extraction using alternative buffers method</p><p>8.3 Example of PCR products using TILLING primers with source genomic DNA from Qiagen kits and low-cost silica method</p><p>8.4 Example of low-cost agarose gel based TILLING assays for the discovery of induced point mutations</p><p>8.5 Example of enzyme activity recorded from weeds compared to crude celery juice extract</p><p>9             Conclusions     </p>