Genome analysis: a practical approach
Methodologies used to analyse the genomes of prokaryotes and eukaryotes are advancing very rapidly as it has been realized that there is a resolution gap between what can be viewed under the microscope and what molecular techniques can resolve. This book presents current, successful strategies for construction of physical maps to identify and analyse the arrangement and function of genes. These techniques, which are applicable to a range of organisms, are being used to move from linked markers to candidate genes in several human monogenic disorders. They are presented in a way that should enable any research or diagnostic laboratory to apply them to their particular systems. Also included are descriptions of new probes and approaches for the localization of human disorders whose analysis has so far eluded the molecular geneticist.
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G S Banting
Microcellmediated Gene Transfer
DNAmediated Gene Transfer
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acrylamide aliquots alleles amplified annealing autoradiography bands blotting buffer Cantor.C.R. centrifuge chromosome chromosome jumping clones concentration containing contig cosmid cycles denaturant denaturing gradient gel described detected DGGE digestion DNA fingerprinting DNA fragment DNA insert DNA molecules DNA polymerase dNTPs double-stranded EcoRl EDTA ethanol ethidium bromide extract Figure filter formamide gel apparatus gel electrophoresis gene genetic genomic DNA hybridization Incubate jumping library labelled lane length ligation litre loading mapping markers medium melting domain method mg/ml microcells microcentrifuge minisatellite mismatch mutations NaCl Natl Notl nucleotides oligonucleotides parallel gel pellet perpendicular gel phage Pharmacia-LKB pipette plasmid plate pmol Preparation primer procedure protocol Re-suspend reaction recipient cells restriction enzyme RFLPs RNA probe RNase RNase cleavage room temperature sequence single-stranded DNA specific stock solution strand supernatant supF template test DNA sample tg/ml transfection transfer Tris-HCl pH tube vector wild-type