Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-74
... transformation of different bacterial strains by plasmids . Bac- teria treated according to the original protocol of Mandel and Higa yield 105–10 ° transformed colonies / μg of supercoiled plasmid DNA . This efficiency can be increased ...
... transformation of different bacterial strains by plasmids . Bac- teria treated according to the original protocol of Mandel and Higa yield 105–10 ° transformed colonies / μg of supercoiled plasmid DNA . This efficiency can be increased ...
Page 1-75
... transformation were achieved with a combination of field strength and pulse length that resulted in 50-75 % cell death . Limited data are available regarding the transformation efficiencies ob- tained with different plasmid DNAs and ...
... transformation were achieved with a combination of field strength and pulse length that resulted in 50-75 % cell death . Limited data are available regarding the transformation efficiencies ob- tained with different plasmid DNAs and ...
Page 1-76
... transformed at frequencies > 5 × 10 ° transformed colonies per microgram of supercoiled plasmid DNA . The maximum frequency of transformation that can be obtained routinely with most other strains of E. coli is approximately five- to ...
... transformed at frequencies > 5 × 10 ° transformed colonies per microgram of supercoiled plasmid DNA . The maximum frequency of transformation that can be obtained routinely with most other strains of E. coli is approximately five- to ...
Contents
DEVELOPMENT OF PLASMID CLONING VECTORS | 1-7 |
Constructing Expression Libraries in Plasmid and Bacteriophage | 1-12 |
Expression Libraries Constructed in Bacteriophage 12 | 1-19 |
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agar plate agarose gel aliquots amber mutations ampicillin antibiotic bacteriophage particles bacteriophage T4 bacteriophage T4 DNA BamHI Bgill buffer carrying cDNA cells cloning coli cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules DNA polymerase double-stranded DNA EcoRI EDTA EDTA pH 8.0 efficiency Escherichia coli ethanol ethidium bromide eukaryotic eukaryotic DNA exonuclease extracts foreign DNA formamide gene genetic HindIII Hindill host hours at 37°C hybridization Incubate infected inserted Kpnl lacZ ligation reaction linear lysogenic method microfuge tube minutes at 4°C nin5 nitrocellulose nitrocellulose filter Nucleic Acids nucleotides packaging pasteur pipette pellet plaques plasmid DNA plasmid vectors polycloning prepared probes protein purified Pvul recA red gam remove replication restriction enzyme RNAase room temperature Sacl Sall sequences Smal solution sterile stored strains strand stuffer supernatant T4 DNA ligase teriophage transfer transformation Tris Cl pH vector DNA vitro volume Xbal µg/ml