Molecular Cloning: A Laboratory Manual, Volume 1

activity agar agarose aliquots allow amount appropriate arms bacterial bacteriophage BamHI buffer carrying cells centrifugation Chapter cloning coli colonies competent concentration construction containing cosmid culture described digestion EcoRI efficiency electrophoresis et al ethanol ethidium ethidium bromide extracts Figure filters final foreign DNA fragments gene genetic genome gradients growth Hindill host hybridization Incubate infected inserted libraries ligase ligation linear method microfuge minutes minutes at 4°C mixture molecules mutations Nucleic Acids obtained original packaging pellet plaques plasmid DNA plate polycloning polymerase possible precipitate prepared presence probes procedure protein purified Pvul reaction recA recombinant Recover region remove replication restriction enzyme resulting room temperature Sall screening segment selection sequences single single-stranded Smal solution step sterile stored strains strand supernatant termini transfer transformation tube usually vector volume yield