Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-7
... VECTORS The development of plasmid vectors has proceeded in three phases : 1. Selectable markers were introduced into plasmids carrying the pMB1 ( or ColE1 ) replicon . The first plasmids used as cloning vectors - pSC101 ( Cohen et al ...
... VECTORS The development of plasmid vectors has proceeded in three phases : 1. Selectable markers were introduced into plasmids carrying the pMB1 ( or ColE1 ) replicon . The first plasmids used as cloning vectors - pSC101 ( Cohen et al ...
Page 2-11
... VECTOR There is no single bacteriophage vector suitable for cloning all DNA fragments . It is therefore necessary to choose carefully among the available vectors for the one best suited to the particular task at hand . Three obvious ...
... VECTOR There is no single bacteriophage vector suitable for cloning all DNA fragments . It is therefore necessary to choose carefully among the available vectors for the one best suited to the particular task at hand . Three obvious ...
Page 4-12
... vectors have also been inserted into a deleted version of pBR322 , creating a family of plasmid vectors ( pUC vectors ) that contain a variety of commonly used cloning sites . These vectors , which are described in detail in Chapter 1 ...
... vectors have also been inserted into a deleted version of pBR322 , creating a family of plasmid vectors ( pUC vectors ) that contain a variety of commonly used cloning sites . These vectors , which are described in detail in Chapter 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