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

Autor Bradley J. Till, Joanna Jankowicz-Cieslak, Owen A. Huynh, Mayada M. Beshir, Robert G. Laport, Bernhard J. Hofinger
en Limba Engleză Hardback – 9 iun 2015
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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Specificații

ISBN-13: 9783319162584
ISBN-10: 3319162586
Pagini: 37
Ilustrații: X, 38 p. 9 illus., 3 illus. in color.
Dimensiuni: 155 x 235 x 15 mm
Greutate: 0.26 kg
Ediția:2015
Editura: Springer International Publishing
Colecția Springer
Locul publicării:Cham, Switzerland

Public țintă

Research

Cuprins

​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.  

Textul de pe ultima copertă

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.

Caracteristici

Open Access provided with a grant from the International Atomic Energy Agency Describes low-cost and easy-to-use assays for the molecular characterization of mutant plant material The experimental protocols do away with the need for complicated procedures, expensive equipment and hazardous chemicals All protocols included have been tried and tested by researchers all over the world Includes supplementary material: sn.pub/extras