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Constructing Expression Libraries in Plasmid and Bacteriophage
Oligonucleotidemediated Mutagenesis 15 51
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acrylamide agar agarose gel aliquots amber mutations bacterial bacteriophage M13 bacteriophage particles bacteriophage T4 DNA Bgill Bgll Bøml buffer carrying CCWGG cDNA cells centrifugation Cl pH cleavage cleaved cloning cohesive termini coli DNA polymerase colonies containing cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules double-stranded DNA EcoRI EDTA efficiency Escherichia coli ethanol ethidium bromide eukaryotic eukaryotic DNA exonuclease exonuclease activity extracts foreign DNA GATC gel electrophoresis gene genetic Gp Cp HindIII host hybridization Incubate infected inserted isoschizomers linear lysogenic method methylase methylation minutes at 4°C nicks nitrocellulose nitrocellulose filters nuclease Nucleic Acids nucleotides packaging pellet phagemids plaques plasmid plasmid DNA plate polycloning prepared probes protein Pvul recA recombinant red gam remove replication restriction enzyme RNAase room temperature SacI sequences single-stranded DNA solution stored strains strand Substrate supernatant T4 DNA ligase template teriophage ug/ml vector DNA vitro volume Xbal