Team:Goettingen/week12-2

V07_16

• Experiment:
  For the chemical retransformation the standard protocol was followed. This time the four E. coli strains DH10B, BL21, DH5alpha and XL1 Blue were transformed in order to subsequently perform swimming assays and compare the accessibility of the different strains.
  
• Observations & Results:
  The retransformation efficient. Numerous colonies had formed at each plate except for the negative control.
  

V07_17

• Experiment:
  In order to test the influence of the different genes on the motility of the distinct strains over night cultures were prepared. On that account it is important to shake the cultures only at 140 rpm to spare the flagella.
  

V07_10

• Experiment:
  In order to clone the new genes into pUC18, all components were digested with EcoRI and XbaI according to the protocol. The restricted plasmid pUC18 was subsequently separated on a gel in order to verify the digestions success and purify the desired fragments. The other digestion products were investigated via preparation of an 1% analytical gel and directly purified using the peqGOLD Gel Extraction Kit (Peqlab) according to the manual as well.
• Observations & Results:
  The digestion was successful. For all PCR products as well as for the vector band of the expected size could be obtained and the fragments can now be used for further cloning.

• Experiment:
  The ligation of the digested genes with the plasmid pUC18 was conducted as described in the protocol.

V07_11

• 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.

V07_12

• Experiment:
  Over night cultures of fliC (DH10B), fliC (Salmonella), motA, motB and yhjH in pUC18 were prepared in order to isolate the plasmids.
  

V07_13

• Experiment:
  Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

• Experiment:
  All vectors were digested with EcoRI and XbaI according to the protocol. The restriction products were subsequently separated on a gel in order to determine successfully ligated plasmids.
  
• Observations & Results:
  Except for fliC (Salmonella) #1 all picked colonies seem to host plasmids containing the aimed insert.