Finding Mutations: The Basics
Many different kinds of mutation can occur in a genome, from large-scale chromosomal rearrangements to small-scale point mutations. This range requires a variety of techniques to detect them; the most difficult is the point mutation, with several alternative techniques available, the suitability of each varying from gene to gene and laboratory to laboratory. This book covers the strategies behind, and methodologies of, mutation detection for the beginner. It will interest graduate students, technicians, and more experienced researchers and clinicians.
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The detection of rearrangements
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acrylamide agarose gel allele allele-specific PCR annealing antisense bands base pair blotting buffer cDNA cent characterized chromosomes clamp cloned codon comb concentration deletion denaturant denaturing gradient gel detection rate DGGE digestion DNA fragments DNA molecules DNA polymerase DNA sequencing DNA strands dot-blot enzyme ethidium bromide exons fluorescence gel apparatus gel electrophoresis gene genomic DNA heteroduplex analysis heterozygous homoduplex homozygous hybridization hydroxylamine insertion isotopic known mutations labelled ligation loading melting domain membrane method microtitre plates migrate mini-sequencing mismatch multiplex mutation detection mutation scanning nonsense mutations nuclease assay Nucleic Acids nucleotide oligo oligonucleotide osmium tetroxide PCR amplification PCR primers PCR product performed perpendicular point mutations polyacrylamide gel polymorphism position pouring probe protein quencher radioactive rearrangements region RNase RNase cleavage screening Section single base single-stranded DNA solution Southern blotting spacers splice SSCP SSCP gels staining target DNA technique temperature tion tissue transcription tube wild-type