Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 3-46
Replica Filters In this method , bacterial colonies grown on nitrocellulose filters
laid on agar plates containing the appropriate antibiotic are frozen in situ
essentially as described by Hanahan and Meselson ( 1980 ) . Approximately 5 x
104 ...
Replica Filters In this method , bacterial colonies grown on nitrocellulose filters
laid on agar plates containing the appropriate antibiotic are frozen in situ
essentially as described by Hanahan and Meselson ( 1980 ) . Approximately 5 x
104 ...
Page 3-52
Preparation of a Transducing Lysate of Packaged Cosmids Because circular
cosmid molecules carried in bacterial cells contain a cos site , they can be
efficiently packaged into bacteriophage à particles . When cosmid - containing
bacteria are ...
Preparation of a Transducing Lysate of Packaged Cosmids Because circular
cosmid molecules carried in bacterial cells contain a cos site , they can be
efficiently packaged into bacteriophage à particles . When cosmid - containing
bacteria are ...
Page 4-48
Suspend a fresh bacterial colony containing pUC118 or pUC119 or their
derivatives in a sterile 15 - ml culture tube containing 2–3 ml of 2 x YT medium .
Add M13K07 to a final concentration of 2 x 10 ' pfu / ml . Incubate for 1.0-1.5
hours at 37 ...
Suspend a fresh bacterial colony containing pUC118 or pUC119 or their
derivatives in a sterile 15 - ml culture tube containing 2–3 ml of 2 x YT medium .
Add M13K07 to a final concentration of 2 x 10 ' pfu / ml . Incubate for 1.0-1.5
hours at 37 ...
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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