Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-92
... filter laid on the surface of a second agar plate . After a period of growth , the colonies are lysed in situ . The master plate is stored at 4 ° C until the results of the screening procedure become available ... Nitrocellulose Filters 1.
... filter laid on the surface of a second agar plate . After a period of growth , the colonies are lysed in situ . The master plate is stored at 4 ° C until the results of the screening procedure become available ... Nitrocellulose Filters 1.
Page 1-94
... nitrocellulose filter and lay it on the master nitrocellulose filter . Take care to prevent air bubbles from becoming trapped between the two filters . This is best done by bending the second filter slightly so that contact is first ...
... nitrocellulose filter and lay it on the master nitrocellulose filter . Take care to prevent air bubbles from becoming trapped between the two filters . This is best done by bending the second filter slightly so that contact is first ...
Page 2-112
... filter prior to hybridization . Because more bacteriophage DNA becomes attached to the nitrocellulose filter , the au- toradiographic signals from positive clones are enhanced ... Nitrocellulose Filters Following Situ Amplification 2.
... filter prior to hybridization . Because more bacteriophage DNA becomes attached to the nitrocellulose filter , the au- toradiographic signals from positive clones are enhanced ... Nitrocellulose Filters Following Situ Amplification 2.
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