Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-53
... DNA is cleaved with a restriction enzyme and joined in vitro to foreign DNA . The resulting recombinant plasmids are then used to transform bacteria . In practice , however , the plasmid vector must be carefully chosen to minimize the ...
... DNA is cleaved with a restriction enzyme and joined in vitro to foreign DNA . The resulting recombinant plasmids are then used to transform bacteria . In practice , however , the plasmid vector must be carefully chosen to minimize the ...
Page 1-54
... Foreign DNA to Plasmid Vectors Termini carried by fragment of foreign DNA Blunt - ended Different protruding termini Identical protruding termini Requirements for cloning high concentrations of DNAs and ligase for maximum efficiency ...
... Foreign DNA to Plasmid Vectors Termini carried by fragment of foreign DNA Blunt - ended Different protruding termini Identical protruding termini Requirements for cloning high concentrations of DNAs and ligase for maximum efficiency ...
Page 4-34
... DNA fragment . If the amount of foreign DNA is limited , use method 2 or 4 . • If the replicative form of the vector DNA has been cleaved with two different restriction enzymes , the fragment of foreign DNA can be inserted in only one ...
... DNA fragment . If the amount of foreign DNA is limited , use method 2 or 4 . • If the replicative form of the vector DNA has been cleaved with two different restriction enzymes , the fragment of foreign DNA can be inserted in only one ...
Contents
Essential Features of Plasmids 1 | 1-3 |
Staininging RNA Before and After Transfer to Nitrocellulose Filters 7 51 | 1-7 |
Index | 1-8 |
Copyright | |
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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