Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 16-38
... ( step 6a , page 16.36 ) or replating the cells in selective medium for isolation of stable transformants ( step 6b , page 16.36 ) . Note If the cells have been proved to survive treatment with glycerol , a glycerol shock ( detailed below ) ...
... ( step 6a , page 16.36 ) or replating the cells in selective medium for isolation of stable transformants ( step 6b , page 16.36 ) . Note If the cells have been proved to survive treatment with glycerol , a glycerol shock ( detailed below ) ...
Page 17-25
... step 8 , page 17.23 , or step 4 , page 17.19 , respectively ) . pAS1 provides a bacteriophage A promoter , PL , and the ribosome - binding site of the bacteriophage à cII gene . Alterna- tively , digest pKK240-11 ( Figure 17.9 ) with ...
... step 8 , page 17.23 , or step 4 , page 17.19 , respectively ) . pAS1 provides a bacteriophage A promoter , PL , and the ribosome - binding site of the bacteriophage à cII gene . Alterna- tively , digest pKK240-11 ( Figure 17.9 ) with ...
Page 17-32
... step 5a and b , and page 17.22 , step 3g - h ) . Cleave the vector at an appropriate downstream site ( EcoRI , SmaI , BamHI ) . 3. Clone the fragment into the vector , transform E. coli strain YK537 , and plate on LB medium containing ...
... step 5a and b , and page 17.22 , step 3g - h ) . Cleave the vector at an appropriate downstream site ( EcoRI , SmaI , BamHI ) . 3. Clone the fragment into the vector , transform E. coli strain YK537 , and plate on LB medium containing ...
Contents
Selectable Markers 1 | 16-7 |
VECTOR SYSTEMS 16 | 16-17 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 16-24 |
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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