Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-40
... minutes at 4 ° C in a Sorvall SS34 rotor ( or its equivalent ) . LiCl precipitates high - molecular - weight RNA . 2. Transfer the supernatant to a fresh 30 - ml Corex tube . Add an equal volume of isopropanol . Mix well . Recover the ...
... minutes at 4 ° C in a Sorvall SS34 rotor ( or its equivalent ) . LiCl precipitates high - molecular - weight RNA . 2. Transfer the supernatant to a fresh 30 - ml Corex tube . Add an equal volume of isopropanol . Mix well . Recover the ...
Page 2-110
... minutes . Transfer the filter into neutralizing solution ( 1.5 M NaCl , 0.5 M Tris Cl ( pH 7.4 ] ) for 5 minutes . Rinse the filter in 2 × SSC , and place it , DNA side up , on paper towels to dry . If any top agarose peels off the ...
... minutes . Transfer the filter into neutralizing solution ( 1.5 M NaCl , 0.5 M Tris Cl ( pH 7.4 ] ) for 5 minutes . Rinse the filter in 2 × SSC , and place it , DNA side up , on paper towels to dry . If any top agarose peels off the ...
Page 4-32
... minutes to 1 hour at room temperature . Stirring for longer periods of time can result in the precipitation of undesired bacterial debris . 3. Collect the precipitate by centrifugation at 10,000g for 20 minutes at 4 ° C . Let the ...
... minutes to 1 hour at room temperature . Stirring for longer periods of time can result in the precipitation of undesired bacterial debris . 3. Collect the precipitate by centrifugation at 10,000g for 20 minutes at 4 ° C . Let 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