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 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 13:16, 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)