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

Inhaltsangabe

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

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