Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 2-102
... hours ) . 3. Induce the lysogen by placing the flasks in a water bath preheated to 45 ° C . Swirl the flasks continuously for 15 minutes . Alternatively , induce the cultures by immersing the flasks in a shaking water bath set at 65 ° C ...
... hours ) . 3. Induce the lysogen by placing the flasks in a water bath preheated to 45 ° C . Swirl the flasks continuously for 15 minutes . Alternatively , induce the cultures by immersing the flasks in a shaking water bath set at 65 ° C ...
Page 4-32
... for 20 minutes at 4 ° C . Let the centrifuge bottle stand in an inverted position for 2-3 minutes to allow the supernatant to drain away from the precipitate . Use Kimwipes to remove the last traces of supernatant from the walls and ...
... for 20 minutes at 4 ° C . Let the centrifuge bottle stand in an inverted position for 2-3 minutes to allow the supernatant to drain away from the precipitate . Use Kimwipes to remove the last traces of supernatant from the walls and ...
Page 6-47
... C on a rotating wheel or rotary platform . Small fragments of DNA ( < 500 bp ) are eluted in 3-4 hours ; larger fragments take 12-16 hours . 6. Centrifuge the sample at 12,000g for 1 minute at 4 ° C in a microfuge . Transfer the ...
... C on a rotating wheel or rotary platform . Small fragments of DNA ( < 500 bp ) are eluted in 3-4 hours ; larger fragments take 12-16 hours . 6. Centrifuge the sample at 12,000g for 1 minute at 4 ° C in a microfuge . Transfer the ...
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