• Preparation of apparatus for the formation of biofilm
Jul 5th Tuesday
Jul.6th
Wednesday
•Receiving primers ordered previously
Jul.7th
Thursday
• Preparation of the aliquot of the primers
• Something wrong with a shaking incubator
Jul.8th
Friday
• Preparation of culture plates for the transformations
Jul.9th
Saturday
• Preparation of culture plates for the transformations
• protocols of transformation
Jul.10th
Sunday
• Several colonies were picked up and cultivated in 5mL LB medium.
•Cryosectioning of biofilm
Week2
Day
Note
Jul.11th Monday
• Set up new LB culture plates with ampicillin and kanamycin
•
·Sterilization of Glycerol and
·Preparation of 25mg/mL kanamycin
•Transformation of the parts mentioned on Jul.9th for the second time
•Observation the sections
Jul 12th Tuesday
• Pick two colonies of each parts and cultivate them in LB medium
•Transformation of three parts(20J,20H.22B from Kit plates of 2011 Distribution ) which are related to the degradation of Cellulose
•·Mini prep to isolate 10I,12I and 22M
•Conservation of 10I,12I,22M and 11P
Jul.13th
Wednesday
• Place the culture plate of 20J,20H and 22B in the fridge.
•Min prep to isolate 13K
•Conservation of 13K
•Restriction digest of the parts(12I,10I,22M,13K) with EcoRI and PstI
•Gel electrophoresis to analyse restriction fragments
•Test the Tm of Primers CP1&CS,NP&NS with 13K. The result of Gel electrophoresis shows that 60.2℃ is the Tm of NS and NP, and 57.4℃ is the Tm of CS and CP1.
Jul.14th
Thursday
• Test the Tm of the YFP,RFP,tetR,Vgb and nirB primers.
The result of Gel electrophoresis shows that the Tm for the primers of Vgb is 54℃ and the Tm for the primers of nirB is 55℃.The RCR of YFP,RFP and tetR failed.
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