Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-21
... culture · Harvesting and lysis of the bacteria • Purification of plasmid DNA Growth of the Bacterial Culture | Plasmids are almost always purified from cultures ( grown in liquid medium containing the appropriate antibiotic ) that have ...
... culture · Harvesting and lysis of the bacteria • Purification of plasmid DNA Growth of the Bacterial Culture | Plasmids are almost always purified from cultures ( grown in liquid medium containing the appropriate antibiotic ) that have ...
Page 1-33
... culture ) with strains of E. coli harboring high - copy - number plasmids carrying the pMB1 or ColE1 replicon . 1. Grow a 30 - ml culture of the bacterial strain carrying the plasmid of interest to late log phase ( i.e. , to an OD 600 ...
... culture ) with strains of E. coli harboring high - copy - number plasmids carrying the pMB1 or ColE1 replicon . 1. Grow a 30 - ml culture of the bacterial strain carrying the plasmid of interest to late log phase ( i.e. , to an OD 600 ...
Page 2-70
... culture of infected bacteria gives the best results ( Blattner et al . 1977 ; Maniatis et al . 1978 ) . 1. Place 100 ml of NZCYM in a 500 - ml flask and inoculate with a single colony of an appropriate bacterial host . Incubate ...
... culture of infected bacteria gives the best results ( Blattner et al . 1977 ; Maniatis et al . 1978 ) . 1. Place 100 ml of NZCYM in a 500 - ml flask and inoculate with a single colony of an appropriate bacterial host . Incubate ...
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 |
Copyright | |
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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