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

by Bradley J. Till, Joanna Jankowicz-Cieslak, Owen A. Huynh, Mayada M. Beshir, Robert G. Laport

Overview

This book offers low-cost and rapid molecular assays for the characterization of mutant plant germplasm. Detailed prools 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.

Product Details

ISBN-13:	9783319386676
Publisher:	Springer International Publishing
Publication date:	07/25/2017
Edition description:	Softcover reprint of the original 1st ed. 2015
Pages:	38
Product dimensions:	6.10(w) x 9.25(h) x 0.00(d)

Table of 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.