Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 3-33
... DNA in the ligation reaction should be greater than 200 μg / ml to favor the formation of mixed concatemers between the arms of the cosmid vector and the eukaryotic DNA . Because of the inability of the vector to self - ligate , the ...
... DNA in the ligation reaction should be greater than 200 μg / ml to favor the formation of mixed concatemers between the arms of the cosmid vector and the eukaryotic DNA . Because of the inability of the vector to self - ligate , the ...
Page 3-38
A Laboratory Manual Joseph Sambrook, E. F. Fritsch, Tom Maniatis. Treatment of Eukaryotic DNA with Alkaline Phosphatase If the supply of eukaryotic DNA is plentiful , prepare it for cloning by partial digestion and fractionation ...
A Laboratory Manual Joseph Sambrook, E. F. Fritsch, Tom Maniatis. Treatment of Eukaryotic DNA with Alkaline Phosphatase If the supply of eukaryotic DNA is plentiful , prepare it for cloning by partial digestion and fractionation ...
Page 3-40
... DNA : eukaryotic DNA . 1. Set up two reaction mixtures as follows : Reaction a fragments of eukaryotic DNA ( 35-45 kb in size ) HindIII / BamHI fragment of pJB8 Sall / BamHI fragment of pJB8 2 με 1 με 1 με 10 ligation buffer ( see page ...
... DNA : eukaryotic DNA . 1. Set up two reaction mixtures as follows : Reaction a fragments of eukaryotic DNA ( 35-45 kb in size ) HindIII / BamHI fragment of pJB8 Sall / BamHI fragment of pJB8 2 με 1 με 1 με 10 ligation buffer ( see page ...
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