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Replication and Incompatibility 1
Constructing Expression Libraries in Plasmid and Bacteriophage
Oligonucleotidemediated Mutagenesis 15 51
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agar plate agarose gel aliquots ampicillin antibiotic bacteriophage particles bacteriophage T4 DNA BamHI buffer carry cDNA cells centrifugation at 12,000g Cl pH cleaved cloning coli containing cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules DNA polymerase double-stranded DNA EcoRI EDTA EDTA pH 8.0 efficiency Escherichia coli ethanol ethidium bromide eukaryotic exonuclease extracts final concentration foreign DNA formamide gel electrophoresis gene genetic gradients Hindlll host hours at 37°C hybridization Incubate infected inserted ligation reaction linear lysogenic method microfuge tube minutes at 4°C minutes at room mixture mutations nitrocellulose nitrocellulose filter Nucleic Acids nucleotides origin of replication packaging pellet phagemid plaques plasmid DNA plasmid vectors polycloning prepared probes protein purified recA recombinant remove replication restriction enzymes RNAase room temperature segment of foreign sequences single-stranded DNA solution step sterile stored strains strand supernatant T4 DNA ligase teriophage transfer transformation vector DNA vitro volume