Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-48
... completely as possible , and store the open tube for a few minutes in an inverted position on a pad of paper towels at room temperature to allow the last traces of ethanol to drain away . 7. Wash the pellet with 70 % ethanol at 4 ° C ...
... completely as possible , and store the open tube for a few minutes in an inverted position on a pad of paper towels at room temperature to allow the last traces of ethanol to drain away . 7. Wash the pellet with 70 % ethanol at 4 ° C ...
Page 5-70
... completely phosphorylated . The reaction at nicks or gaps in double- stranded DNA is less efficient than for single - stranded termini ; however , with sufficient concentrations of ATP and enzyme , gaps can be completely phosphorylated ...
... completely phosphorylated . The reaction at nicks or gaps in double- stranded DNA is less efficient than for single - stranded termini ; however , with sufficient concentrations of ATP and enzyme , gaps can be completely phosphorylated ...
Page 5-72
... completely at the end of dephosphorylation reactions . Proteinase K is used to digest CIP , which must be completely removed if subsequent ligations are to work efficiently . An alternative method is to inactivate the CIP by heating to ...
... completely at the end of dephosphorylation reactions . Proteinase K is used to digest CIP , which must be completely removed if subsequent ligations are to work efficiently . An alternative method is to inactivate the CIP by heating 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