Difference between revisions of "PGP1331"
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'''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 | + | * skip the Stopp-Codon of the gene in the reverse 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 13:23, 24 June 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 reverse primer
- take care that the integration is in-frame
Antibody
- dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)
