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Low-Cost Methods for Molecular Characterization of Mutant Plants [electronic resource] : Tissue Desiccation, DNA Extraction and Mutation Discovery: Protocols / by Bradley J. Till, Joanna Jankowicz-Cieslak, Owen A. Huynh, Mayada M. Beshir, Robert G. Laport, Bernhard J. Hofinger.

By: Contributor(s): Material type: TextTextPublisher: Cham : Springer International Publishing : Imprint: Springer, 2015Edition: 1st ed. 2015Description: X, 35 p. 9 illus., 3 illus. in color. online resourceContent type:
  • text
Media type:
  • computer
Carrier type:
  • online resource
ISBN:
  • 9783319162591
Subject(s): Additional physical formats: Printed edition:: No title; Printed edition:: No title; Printed edition:: No titleDDC classification:
  • 631.52 23
  • 660.6 23
LOC classification:
  • SB123-123.5
  • S494.5.B563
Online resources:
Contents:
Introduction -- Health and Safety Considerations -- Sample Collection and Storage -- Low-Cost DNA Extraction -- PCR Amplification for Low-Cost Mutation Discovery -- Enzymatic Mismatch Cleavage and Agarose Gel Evaluation of Samples -- Alternative Enzymology for Mismatch Cleavage for TILLING and Ecotilling: Extraction of Enzymes form Common Weedy Plants -- Example Data -- Conclusions.  .
In: Springer eBooksSummary: This book offers low-cost and rapid molecular assays for the characterization of mutant plant germplasm. Detailed protocols are provided for the desiccation of plant tissues; the extraction of high-quality DNA for downstream applications; the extraction of single-strand-specific nucleases for single nucleotide polymorphism; and small insertion/deletion discovery using standard agarose gel electrophoresis. The methods described can be applied in any laboratory equipped for basic molecular biology and do away with the need for expensive freezers and toxic organic compounds. With the appropriate validation of sample quality and longevity, they can provide sufficient DNA for a variety of molecular applications, such as marker studies and TILLING, at approximately one tenth of the cost per sample when compared to commercial kits.
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Introduction -- Health and Safety Considerations -- Sample Collection and Storage -- Low-Cost DNA Extraction -- PCR Amplification for Low-Cost Mutation Discovery -- Enzymatic Mismatch Cleavage and Agarose Gel Evaluation of Samples -- Alternative Enzymology for Mismatch Cleavage for TILLING and Ecotilling: Extraction of Enzymes form Common Weedy Plants -- Example Data -- Conclusions.  .

Open Access

This book offers low-cost and rapid molecular assays for the characterization of mutant plant germplasm. Detailed protocols are provided for the desiccation of plant tissues; the extraction of high-quality DNA for downstream applications; the extraction of single-strand-specific nucleases for single nucleotide polymorphism; and small insertion/deletion discovery using standard agarose gel electrophoresis. The methods described can be applied in any laboratory equipped for basic molecular biology and do away with the need for expensive freezers and toxic organic compounds. With the appropriate validation of sample quality and longevity, they can provide sufficient DNA for a variety of molecular applications, such as marker studies and TILLING, at approximately one tenth of the cost per sample when compared to commercial kits.

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