Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-29
... tube in a boiling - water bath for exactly 40 seconds . 4. Centrifuge the bacterial lysate at 12,000g for 10 minutes at room temperature in a microfuge . 5. Remove the pellet of bacterial debris from the microfuge tube with a sterile ...
... tube in a boiling - water bath for exactly 40 seconds . 4. Centrifuge the bacterial lysate at 12,000g for 10 minutes at room temperature in a microfuge . 5. Remove the pellet of bacterial debris from the microfuge tube with a sterile ...
Page 1-44
... tube . This speeds the formation of the CsCl gradient and allows the centrifugation time to be reduced to 6 hours . 1. Prepare a solution of CsCl ( p = 1.47 g / ml ) by adding 125 g of CsCl to 167 ml of TE ( pH 8.0 ) . 2. Add 8 ml of ...
... tube . This speeds the formation of the CsCl gradient and allows the centrifugation time to be reduced to 6 hours . 1. Prepare a solution of CsCl ( p = 1.47 g / ml ) by adding 125 g of CsCl to 167 ml of TE ( pH 8.0 ) . 2. Add 8 ml of ...
Page 7-21
... tube just above the level of the remaining fluid . Place the bottom of the tube on a pad of Kimwipes , and carefully withdraw the fluid with an automatic pipettor . Invert the tube and allow any remaining fluid to drain into the pad of ...
... tube just above the level of the remaining fluid . Place the bottom of the tube on a pad of Kimwipes , and carefully withdraw the fluid with an automatic pipettor . Invert the tube and allow any remaining fluid to drain into the pad of ...
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