Difference between revisions of "PGP1331"
| Line 3: | Line 3: | ||
'''General comments''' | '''General comments''' | ||
| − | Integrative | + | Integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor |
| − | *E.coli: multicopy plasmid, select for ampicillin | + | * ''E.coli'': multicopy plasmid, select for ampicillin |
| − | *B. subtilis: no ori, select for spectinomycin | + | * ''B. subtilis'': no ori, select for spectinomycin |
'''Hints for the usage''' | '''Hints for the usage''' | ||
| − | *amplify the last 600bp of your gene of interest, | + | * amplify the last 600bp of your gene of interest, |
| − | *skip the Stopp-Codon of the gene in the revers primer | + | * skip the Stopp-Codon of the gene in the revers primer |
| − | *take care that the integration is in-frame | + | * take care that the integration is in-frame |
'''Antibody''' | '''Antibody''' | ||
*dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it) | *dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it) | ||
Revision as of 13:22, 16 March 2010
General comments
Integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor
- E.coli: multicopy plasmid, select for ampicillin
- B. subtilis: no ori, select for spectinomycin
Hints for the usage
- amplify the last 600bp of your gene of interest,
- skip the Stopp-Codon of the gene in the revers primer
- take care that the integration is in-frame
Antibody
- dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)
