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 1-59
... DNA ligase and of foreign and plasmid DNAs . In addition , the efficiency of this type of reaction is often increased by addition of low concentrations of substances such as polyethylene glycol ( see page 1.70 ) . The ... Foreign DNA 1.
... DNA ligase and of foreign and plasmid DNAs . In addition , the efficiency of this type of reaction is often increased by addition of low concentrations of substances such as polyethylene glycol ( see page 1.70 ) . The ... Foreign DNA 1.
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