Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-78
... aliquots , and store the aliquots in tissue - culture flasks ( Corning 25100 or equivalent ) at 4 ° C . The pH of the solution should be between 6.0 and 6.1 . a b Rubidium chloride ( 100 mM ) was included in the original recipe for TFB ...
... aliquots , and store the aliquots in tissue - culture flasks ( Corning 25100 or equivalent ) at 4 ° C . The pH of the solution should be between 6.0 and 6.1 . a b Rubidium chloride ( 100 mM ) was included in the original recipe for TFB ...
Page 1-79
... aliquots in sterile tubes . The tubes should be closed tightly and stored at -70 ° C . Each aliquot should be used only once and discarded . For preparation of 1 M potassium acetate ( pH 7.5 ) , see Table 1.4 . ii . Add an additional ...
... aliquots in sterile tubes . The tubes should be closed tightly and stored at -70 ° C . Each aliquot should be used only once and discarded . For preparation of 1 M potassium acetate ( pH 7.5 ) , see Table 1.4 . ii . Add an additional ...
Page 1-80
... aliquots of the competent - cell suspension will be more than adequate . However , when large numbers of transformed colonies are required ( e.g. , when constructing cDNA libraries ) , larger aliquots may be needed . iv . When needed ...
... aliquots of the competent - cell suspension will be more than adequate . However , when large numbers of transformed colonies are required ( e.g. , when constructing cDNA libraries ) , larger aliquots may be needed . iv . When needed ...
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