IGEM:Groningen/Notebook/iGEM 2011/2011/07/06

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6th of July


* Colony PCR of 6 constructs!
HybB promotor in pSB1CR
HybB promotor in RBS GFP ampicillin resistance vector
PcI promotor in RBS GFP ampicillin resistance vector
PlasI promotor in RBS GFP ampicillin resistance vector
LasI-LVA in ampicillin resistance with double terminator vector
cI-LVA in ampicillin resistance with double terminator vector

Colony PCR protocol:
Media: Taq PCR mastermix
Preheated Lid: 111° C
Denaturation: 95 ° C for ten minutes
Cycle 33×
Denaturation: 95° C for 30 seconds
Annealing: 60° C for 30 seconds
Extension: 72° C for 1 minute
Final extension: 72° C for ten minutes
Store at 4 ° C (infinite)