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This book is open access under a CC BY-NC 2.5 license. This book
offers 19 detailed protocols on the use of induced mutations in
crop breeding and functional genomics studies, which cover topics
including chemical and physical mutagenesis, phenotypic screening
methods, traditional TILLING and TILLING by sequencing, doubled
haploidy, targeted genome editing, and low-cost methods for the
molecular characterization of mutant plants that are suitable for
laboratories in developing countries. The collection of protocols
equips users with the techniques they need in order to start a
program on mutation breeding or functional genomics using both
forward and reverse-genetic approaches. Methods are provided for
seed and vegetatively propagated crops (e.g. banana, barley,
cassava, jatropha, rice) and can be adapted for use in other
species.
This book is open access under a CC BY-NC 2.5 license. This book
offers 19 detailed protocols on the use of induced mutations in
crop breeding and functional genomics studies, which cover topics
including chemical and physical mutagenesis, phenotypic screening
methods, traditional TILLING and TILLING by sequencing, doubled
haploidy, targeted genome editing, and low-cost methods for the
molecular characterization of mutant plants that are suitable for
laboratories in developing countries. The collection of protocols
equips users with the techniques they need in order to start a
program on mutation breeding or functional genomics using both
forward and reverse-genetic approaches. Methods are provided for
seed and vegetatively propagated crops (e.g. banana, barley,
cassava, jatropha, rice) and can be adapted for use in other
species.
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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