Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 49
Incubate the suspension for 5 minutes at 4°C . Lysozyme will not work efficiently if
the pH of the solution ... 0 ) , and incubate the suspension for 15 minutes at 37°C .
The bacterial cell wall is digested during this incubation . The resulting ...
Incubate the suspension for 5 minutes at 4°C . Lysozyme will not work efficiently if
the pH of the solution ... 0 ) , and incubate the suspension for 15 minutes at 37°C .
The bacterial cell wall is digested during this incubation . The resulting ...
Page 19
Incubate the reaction for 30 minutes at 37°C . c . Heat the reaction for 5 minutes
at 68°C to inactivate the bacteriophage T4 polynucleotide kinase . Place on dry
ice and store at - 20°C . 3 . Hybridize the primer to template bacteriophage M13 ...
Incubate the reaction for 30 minutes at 37°C . c . Heat the reaction for 5 minutes
at 68°C to inactivate the bacteriophage T4 polynucleotide kinase . Place on dry
ice and store at - 20°C . 3 . Hybridize the primer to template bacteriophage M13 ...
Page 56
Incubate the gel for 30 minutes at room temperature with gentle shaking . 3 .
Repeat step 2 . 4 . Discard the ethanol and add 10 gel volumes of deionized
water . Incubate the gel for 10 minutes at room temperature with gentle shaking .
5 .
Incubate the gel for 30 minutes at room temperature with gentle shaking . 3 .
Repeat step 2 . 4 . Discard the ethanol and add 10 gel volumes of deionized
water . Incubate the gel for 10 minutes at room temperature with gentle shaking .
5 .
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Contents
BOOK | 3 |
Identification and Analysis of Recombinants 2 108 | 30 |
TRANSFECTION MEDIATED BY DEAEDEXTRAN 16 | 41 |
Copyright | |
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Common terms and phrases
acetate activity added Adjust aliquots allow amount antibody antigen Appendix appropriate approximately aqueous assay bacterial bacteriophage base binding buffer carried cells centrifugation Chapter chromatography Cl pH cloned coli column complete concentration containing culture deionized water described desired detected dilution Dissolve EDTA efficiency electrophoresis enzyme et al ethanol expression extract Figure filter final fragment gene growth hybridization Incubate levels ligation mammalian cells medium method mg/ml microfuge tube minutes mixture mRNAs nucleic acid pellet phase phosphate Place plasmid plate polymerase precipitation prepared procedure Products promoter protein radioactive radiolabeled reaction remove restriction room temperature sample screen selection sequence sodium solution specific step sterile stored strain TABLE Telephone termini tion transcription transfection transfer transformation Tris ug/ml usually vectors volume wash western blotting ОН