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

• Author: Till, Bradley J.
• TeilnehmendeR: Jankowicz-Cieslak, Joanna.; Huynh, Owen A.; Beshir, Mayada M.; Laport, Robert G.; Hofinger, Bernhard J.
• Place / Publishing House: Cham : : Springer International Publishing AG,, 2015.; ©2015.
• Year of Publication: 2015
• Edition: 1st ed.
• Language: English
• Physical Description: 1 online resource (42 pages)

Table of Contents

• Intro
• Foreword
• Acknowledgements
• Contents
• Chapter 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
• Chapter 2: Health and Safety Considerations
• 2.1 Guidelines
• 2.2 Preparation of a Home-Made Chemical Spill Kit
• Reference
• Chapter 3: Sample Collection and Storage
• 3.1 Background
• 3.2 Materials
• 3.3 Methods
• References
• Chapter 4: Low-Cost DNA Extraction
• 4.1 Materials
• 4.2 Methods
• 4.2.1 Preparation of Silica Powder DNA Binding Solution
• 4.2.2 Low-Cost Extraction of Genomic DNA
• 4.3 Alternative Buffers for DNA Extraction
• Reference
• Chapter 5: PCR Amplification for Low-Cost Mutation Discovery
• 5.1 Materials
• 5.2 Methods
• References
• Chapter 6: Enzymatic Mismatch Cleavage and Agarose Gel Evaluation of Samples
• 6.1 Materials
• 6.2 Methods
• Reference
• Chapter 7: Alternative Enzymology for Mismatch Cleavage for TILLING and Ecotilling: Extraction of Enzymes from Common Weedy Pl...
• 7.1 Materials
• 7.2 Methods
• 7.2.1 Enzyme Extraction
• 7.2.2 Concentration of Enzymes Using Amicon Ultra 10 kDa MWCO Centrifugal Filter Devices (for 0.5 ml Starting Volume
• in 1.5-m...
• 7.2.3 Test of Mismatch Cleavage Activity
• Reference
• Chapter 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 Method Using Alternative Buffers
• 8.2.1 Summary
• 8.3 Example of PCR Products Using TILLING Primers with Source Genomic DNA from a Commercial Kit 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 Recovered from Weeds Compared to Crude Celery Juice Extract
• 8.5.1 Summary
• References
• Chapter 9: Conclusions.