Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 121
Many of the currently used plasmid vectors ( e.g. , the PUC series ) replicate to
such a high copy number that they can be purified in large yield from cultures that
have simply been grown to late log phase in standard LB medium . In these ...
Many of the currently used plasmid vectors ( e.g. , the PUC series ) replicate to
such a high copy number that they can be purified in large yield from cultures that
have simply been grown to late log phase in standard LB medium . In these ...
Page 174
Bacteria treated according to the original protocol of Mandel and Higa yield 108–
108 transformed colonies / ug of supercoiled plasmid DNA . This efficiency can
be increased 100- to 1000 - fold by exposing improved strains of E. coli ( Kushner
...
Bacteria treated according to the original protocol of Mandel and Higa yield 108–
108 transformed colonies / ug of supercoiled plasmid DNA . This efficiency can
be increased 100- to 1000 - fold by exposing improved strains of E. coli ( Kushner
...
Page 4-49
Screening Colonies by Superinfection Several colonies can be screened
simultaneously for their ability to yield single - stranded DNA after superinfection .
1. Suspend a fresh bacterial colony containing pUC118 or pUC119 or their
derivatives ...
Screening Colonies by Superinfection Several colonies can be screened
simultaneously for their ability to yield single - stranded DNA after superinfection .
1. Suspend a fresh bacterial colony containing pUC118 or pUC119 or their
derivatives ...
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Common terms and phrases
activity agar agarose aliquots allow amount appropriate arms bacterial bacteriophage BamHI buffer carrying cells centrifugation Chapter cloning coli colonies completely 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 growth Hindi host hybridization Incubate infected inserted libraries ligase ligation linear method microfuge minutes minutes at 4°C mixture molecules mutations nucleic acids obtained original packaging particles pellet plaques plasmid DNA plate polycloning polymerase possible precipitate prepared presence probes problem procedure promoter protein purified reaction recombinant Recover region remove replication restriction enzyme room temperature Sall segment selection sequences single single-stranded Smal solution step sterile stored strains strand supernatant termini transfer transformation tube usually vector volume Xbal yield