Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page xxxi
... LINKERS INTO PROTEIN - CODING SEQUENCES 15.85 Designing the Hexameric Linker 15.86 Insertion of Hexameric Linkers 15.88 CREATING MANY MUTATIONS IN A DEFINED SEGMENT OF DNA 15.95 Use of Degenerate Pools of Mutagenic Oligonucleotides ...
... LINKERS INTO PROTEIN - CODING SEQUENCES 15.85 Designing the Hexameric Linker 15.86 Insertion of Hexameric Linkers 15.88 CREATING MANY MUTATIONS IN A DEFINED SEGMENT OF DNA 15.95 Use of Degenerate Pools of Mutagenic Oligonucleotides ...
Page xxxviii
... Linkers F.8 Ligation of Phosphorylated Linkers to Blunt - ended Target Fragments F.9 Appendix G : List of Suppliers References Molecular Cloning Plasmid Vectors Bacterial plasmids are double - stranded xxxviii Contents.
... Linkers F.8 Ligation of Phosphorylated Linkers to Blunt - ended Target Fragments F.9 Appendix G : List of Suppliers References Molecular Cloning Plasmid Vectors Bacterial plasmids are double - stranded xxxviii Contents.
Page 1-28
... linker insertion , 15.5 of hexameric linkers into protein coding sequence , 15.85-15.94 by partial digestion with DNAase I , 15.5-15.7 using restriction enzymes , 15.8– 15.13 linker - scanning , 15.32-15.50 loop - out mutagenesis , 15.3 ...
... linker insertion , 15.5 of hexameric linkers into protein coding sequence , 15.85-15.94 by partial digestion with DNAase I , 15.5-15.7 using restriction enzymes , 15.8– 15.13 linker - scanning , 15.32-15.50 loop - out mutagenesis , 15.3 ...
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