Low-Cost Methods for Molecular Characterization of Mutant Plants: Tissue Desiccation, DNA Extraction and Mutation Discovery: Protocols (Hardcover, 2015)

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