The cultures from shift 2 had similar results as shift one. However, the GFP signal measured from the LigA construct was stronger. Thus, cells definitely need to grow before undergoing irradiation.
The dilutions and the shift one cultures put overnight did not show any significant results.
was miniprepped and nanodropped
Part
concentration (ng/μl)
260/280 λ
(A)
238,9
1,95
(B)
248,8
1,91
After nanodropping, was restricted with SpeI and PstI, while was cut with PstI and XbaI.
A gel electrophoresis and extraction was performed. This showed an unexpected 800 bp signal at the lanes with . This is in fact due to the vector in which this part is ligated. contains a reporter gene, RFP. Thus, in combination with the promotor, a construct is formed that can be used to measure activity of this promotor. Since the promotor is flanked by standard assembly restriction sites we can replace the current promotor () by any other promotor, for instance .
DB3.1, and LigA were ented in liquid culture and grown overnight to make glycerol stocks and competent cells
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