Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-66
... vector DNA to achieve a j : i ratio of > 1 and < 3 . For a plasmid the size of pUC18 , this means that the ligation reaction should contain between 20 μg and 60 μg of vector DNA per milliliter . • An equal or slightly greater ...
... vector DNA to achieve a j : i ratio of > 1 and < 3 . For a plasmid the size of pUC18 , this means that the ligation reaction should contain between 20 μg and 60 μg of vector DNA per milliliter . • An equal or slightly greater ...
Page 2-11
... VECTOR There is no single bacteriophage A vector suitable for cloning all DNA fragments . It is therefore necessary to choose carefully among the available vectors for the one best suited to the particular task at hand . Three obvious ...
... VECTOR There is no single bacteriophage A vector suitable for cloning all DNA fragments . It is therefore necessary to choose carefully among the available vectors for the one best suited to the particular task at hand . Three obvious ...
Page 2-82
... vector DNA by cleavage with appropriate restriction en- zyme ( s ) . 2. Ligation of the digested vector with fragments of foreign DNA having termini compatible with ... Vectors Cloning in Bacteriophage λ 2 82 PREPARATION OF VECTOR DNA 2 82.
... vector DNA by cleavage with appropriate restriction en- zyme ( s ) . 2. Ligation of the digested vector with fragments of foreign DNA having termini compatible with ... Vectors Cloning in Bacteriophage λ 2 82 PREPARATION OF VECTOR DNA 2 82.
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