Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-34
... ( pH 8.0 ) 1 mM EDTA ( pH 8.0 ) 3. Collect the bacterial cells by centrifugation as described in step 1 . LYSIS BY BOILING This method ( adapted from Holmes and Quigley 1981 ) is designed to be used in conjunction with a subsequent ...
... ( pH 8.0 ) 1 mM EDTA ( pH 8.0 ) 3. Collect the bacterial cells by centrifugation as described in step 1 . LYSIS BY BOILING This method ( adapted from Holmes and Quigley 1981 ) is designed to be used in conjunction with a subsequent ...
Page 6-55
... EDTA ( pH 8.0 ) , 0.01 M Tris Cl ( pH 7.6 ) . 2. Resuspend the cells at a concentration of 3 × 1010 cells / ml in 0.05 M EDTA ( pH 8.0 ) at 0 ° C . 3. Prepare an equal volume of low - melting - temperature agarose ( 1 % ) in L buffer ...
... EDTA ( pH 8.0 ) , 0.01 M Tris Cl ( pH 7.6 ) . 2. Resuspend the cells at a concentration of 3 × 1010 cells / ml in 0.05 M EDTA ( pH 8.0 ) at 0 ° C . 3. Prepare an equal volume of low - melting - temperature agarose ( 1 % ) in L buffer ...
Page 7-24
... 8 M guanidine HCl ( M = 95.6 ) 0.1 M sodium acetate ( pH 7.0 ) 1 mм dithiothreitol 20 mM EDTA ( pH 8.0 ) Add 191 g of guanidine HCl to 8.35 ml of 3 м sodium acetate ( pH 7.0 ) , 1.25 ml of 0.2 м dithiothreitol , and 10 ml of 0.5 M EDTA ( pH ...
... 8 M guanidine HCl ( M = 95.6 ) 0.1 M sodium acetate ( pH 7.0 ) 1 mм dithiothreitol 20 mM EDTA ( pH 8.0 ) Add 191 g of guanidine HCl to 8.35 ml of 3 м sodium acetate ( pH 7.0 ) , 1.25 ml of 0.2 м dithiothreitol , and 10 ml of 0.5 M EDTA ( pH ...
Contents
BOOK | 1-2 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 1-7 |
Index | 1-8 |
Copyright | |
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agar agarose gel aliquots amber mutations bacteria bacteriophage M13 bacteriophage particles bacteriophage T4 bacteriophage T4 DNA BamHI Bgill buffer cDNA cells centrifugation at 12,000g cleaved cloning coli DNA polymerase colonies containing cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules DNA polymerase double-stranded DNA EcoRI EDTA efficiency Escherichia coli ethanol ethidium bromide eukaryotic eukaryotic DNA exonuclease extracts foreign DNA gel electrophoresis gene genetic Gp Cp HindIII Hindill host hybridization Incubate infected inserted intergenic region Kpnl lacZ linear lysogenic method methylase methylation microfuge tube minutes at 4°C nin5 nitrocellulose nucleotides origin of replication packaging pellet phagemids plaques plasmid plasmid DNA plate polycloning prepared protein Pvul recA recombinant red gam remove replication restriction enzyme room temperature Sacl Sall single-stranded DNA Smal solution sterile stored strains strand supernatant T4 DNA ligase teriophage Transfer Tris Cl pH vector DNA vitro volume Xbal µg/ml