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Replication and Incompatibility 1
Constructing Expression Libraries in Plasmid and Bacteriophage
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agarose gel aliquots ampicillin bacterial bacteriophage M13 bacteriophage particles bacteriophage T4 DNA buffer carry CCWGG cDNA cells centrifugation at 12,000g Cl pH cleavage cleaved cloning cohesive termini coli DNA polymerase colonies containing cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules DNA polymerase double-stranded DNA EcoRI EDTA EDTA pH 8.0 efficiency ethanol ethidium bromide eukaryotic eukaryotic DNA exonuclease exonuclease activity extracts final concentration foreign DNA GATC gel electrophoresis gene genomic DNA HindIII hybridization Incubate infected inserted isoschizomers ligation reaction method methylase methylation microfuge tube minutes at 4°C mixture nitrocellulose nitrocellulose filter nuclease Nucleic Acids nucleotides origin of replication packaging pellet phagemids plaques plasmid plasmid DNA polycloning prepared probes protein radiolabeled recA recombinant cosmids remove replication restriction enzyme RNAase room temperature sequences single-stranded DNA solution sterile stored strains strand Substrate supernatant T4 DNA ligase template teriophage transfer ug/ml vector DNA vitro volume Xbal