Creation of the cro-box testing devices

Kamil

Tasks:

• Ligation of the device with two variants of the cro-box
• Bacteria transformation
• I14018 brick resurrection
• R0010 brick multiplication

Methods:

• The ligation mix was prepared as follows (same for both variants):

  20μl purified cro-box plasmid 
  20mu;l purified reporter device
  5μl ligation buffer (Fermentas)
  1μl T4 Ligase (Fermentas)
  4μl H2O

• A fresh bach o chemocompetent bacteria was transformed with the ligation mixtures and incubated overnight on agarose plates supplemented with ampicillin.
• A fresh bach o chemocompetent bacteria was transformed with the I14018 brick solution and incubated overnight on agarose plates supplemented with kanamycin.
• Bacterial culture carrying the Roo10 brick was resuspended in 50ml of liquid LB medium suplemented with ampicillin and incubated overnight.