Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 18-73
... Enzyme - coupled secondary reagents 1. Transfer the nitrocellulose filter from the final wash in PBS ( step 3 , page 18.71 ) to a tray containing 200 ml of 150 mм NaCl , 50 mM Tris Cl ( pH 7.5 ) . Incubate the filter for 10 minutes at ...
... Enzyme - coupled secondary reagents 1. Transfer the nitrocellulose filter from the final wash in PBS ( step 3 , page 18.71 ) to a tray containing 200 ml of 150 mм NaCl , 50 mM Tris Cl ( pH 7.5 ) . Incubate the filter for 10 minutes at ...
Page F-4
... enzymes . The exonuclease reaction can therefore be carried out by adding the polymerase directly to the digestion mixture together with high concentrations of the four dNTPs . The enzyme removes protruding 3 ' nucleotides until it ...
... enzymes . The exonuclease reaction can therefore be carried out by adding the polymerase directly to the digestion mixture together with high concentrations of the four dNTPs . The enzyme removes protruding 3 ' nucleotides until it ...
Page F-10
... enzyme buffer appropriate restriction enzyme 70 μ . 10 με 20-50 units Mix , and then incubate the reaction for 4 hours at the optimal tempera- ture for the restriction enzyme . 4. Add an additional 10 units of the restriction enzyme ...
... enzyme buffer appropriate restriction enzyme 70 μ . 10 με 20-50 units Mix , and then incubate the reaction for 4 hours at the optimal tempera- ture for the restriction enzyme . 4. Add an additional 10 units of the restriction enzyme ...
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Common terms and phrases
Acad acetate acrylamide activity agarose aliquots amount antibody antigen aqueous phase assay autoclaving autoradiographic bacterial bacteriophage T4 DNA BamHI Biol blunt-ended cDNA cell lines centrifugation chloramphenicol chromatography cloned gene coli DNA polymerase column containing culture Dissolve dithiothreitol DNA ligase DNA polymerase dNTPs double-stranded EDTA EDTA pH 8.0 efficiency electrophoresis eluted encoding Escherichia coli ethanol ethidium bromide eukaryotic expression vectors extract gel-loading buffer H₂O hybridization Incubate intensifying screen ligation linkers mammalian cells medium method mg/ml microfuge tube mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid nucleotides OH OH P.O. Box pellet phosphate pipette plasmid PMSF polycloning precipitation prepared Proc promoter radioactive radiolabeled reaction remove restriction enzyme room temperature sample sequences sodium sterile stock solution stored strain supernatant target protein Telephone termini tion transcription transfection transfer Tris Cl pH vectors washed western blotting µg/ml