Team:Goettingen/week6-2

V06_04

• Experiment:
  For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:
  20E - #2
  20E - #3
  18M - #2
  18M - #3
  18C - #1
  18C - #2
  
• Observations & Results:
  The retransformation was successful since all plates showed numeours colonoes except the negative control.
  

V06_05

• Experiment:
  In order to clone flhDC behind further promoters, the purified PCR product was digested with XbaI and PstI, whereas the BioBrick plasmids were cut with SpeI and PstI. The digestion was performed as described in the protocol. This time the follwing promoters were used:
  J23113 - 20G - x21
  J23109 - 2G - x106
  J23114 - 20I - x256
  J23106 - 18O - x1185
  J23104 - 18K - x 1831
  

• Experiment:
  The ligation of the digested flhDC with the BioBrick plasmids was conducted as described in the protocol.
  

V06_06

• Experiment:
  For the chemical transformation the standard protocol was followed.
• Observations & Results:
  The transformation was successful since all plates showed numeours colonoes except the negative control. However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected.

V06_07

• Experiment:
  Over night cultures were prepapred of three colonies of each constructs in order to isolate the plasmids.
  

V06_08

• Experiment:
  Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
• 
  V06_08_2 Test digestion of the new flhDC-promoter constructs
  
  • Experiment:
    In order to prove correct insertion of flhDC a test digestion was performed using SpeI and XbaI according to the protocol.
    
  • Observations & Results:
    For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.