Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-52
... minutes at room temperature in a Beckman TJ6 cen- trifuge ( or its equivalent ) . Remove the supernatant by aspiration . 4. Flick the side of the tube with a finger until the pellet is loosened , and then add 0.5 ml of 50 % polyethylene ...
... minutes at room temperature in a Beckman TJ6 cen- trifuge ( or its equivalent ) . Remove the supernatant by aspiration . 4. Flick the side of the tube with a finger until the pellet is loosened , and then add 0.5 ml of 50 % polyethylene ...
Page 17-41
... hour at room temperature . 4 3. Add this solution to 9 volumes of 50 mm KH2PO ( pH 10.7 ) , 1 mм EDTA ( pH 8.0 ) , 50 mM NaCl , and store for 30 minutes at room temperature . Maintain the pH at 10.7 with KOH . Caution : Handle ...
... hour at room temperature . 4 3. Add this solution to 9 volumes of 50 mm KH2PO ( pH 10.7 ) , 1 mм EDTA ( pH 8.0 ) , 50 mM NaCl , and store for 30 minutes at room temperature . Maintain the pH at 10.7 with KOH . Caution : Handle ...
Page 18-56
... 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. Repeat step 4 twice ...
... 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. Repeat step 4 twice ...
Contents
Selectable Markers 1 | 16-7 |
VECTOR SYSTEMS 16 | 16-17 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 16-24 |
Copyright | |
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Acad acetate acrylamide activity Adjust the volume agarose aliquots amount antibody antigen antisera aqueous assay autoclaving bacterial bacteriophage T4 BamHI Biol calcium cDNA cell lines centrifugation chloramphenicol cloned gene column containing culture deionized H₂O dhfr dilution Dissolve dithiothreitol DNA polymerase EDTA EDTA pH 8.0 efficiency elution encoding Escherichia coli ethanol ethidium bromide eukaryotic extract fragment glycerol hybridization immunoglobulin immunoprecipitation Incubate ligation lysate mammalian cells medium method mg/ml microfuge tube minutes at 4°C minutes at room mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid pellet phosphate plasmid plate PMSF polyacrylamide polyclonal precipitate prepared promoter protoplasts radioactive radiolabeled reaction reagent recombinant remove restriction enzyme Resuspend room temperature sample SDS gel-loading buffer SDS-polyacrylamide gel sequences serum sodium sterile stock solution stored strain supernatant target protein Telephone termini transcription transfection transfer transient expression Tris Cl pH wash western blotting µg/ml