21.7.2023

Events:Visited the laboratory, have the basic concept of the layout of the laboratory. Learned how to prepare LB solution to breed bacteria. Use the LB prepared by students to feed and increase the number of E.coli for the experiment next day.

 

22.7.2023

Events:Extract plasmid base from some E.coli prepared yesterday and detect the concentration, this procedure is failed as the concentration is very low. Put the lasted E.coli to LB in the clean bench to ensure the bacteria in the solution only have E.coli, these LB-bacteria solution is for next day’s experiment.

 

23.7.2023

Events:Extract plasmid base and target fragment from some E.coli prepared yesterday and detect the concentration, this procedure is seccessful as the concentration is at normal value. The digestion is then used to get the target fragment and plasmid base and then use agarose gel electrophoresis to detect whether the digestion is seccessed .

 

24.7.2023

Events: Cut the agarose gel containing target fragment and extract the fragments out, detect the concentration. The extraction of fragments in the agarose gel is successful as the concentration is at normal value. Use enzymic linkage to construct a hole plasmid with target fragment with plasmid base. Move the plasmid in to competent cell and feed the target-plasmid E.coli in the LB solution with Kana.

 

25.7.2023

Events:The E.coil cannot grow in LB-Kana solution. The transform was failed. Attract the plasmid and target fragment from the lasted E.coli and detect the concentration. The extraction is successful as the concentration is at normal level. The digestion is succeccful as the concentration is at normal level. The agarose gel electrophoresis is successful as the graph under UV lamp shows two strips. The part of gel contain fragments are then recycled and detected the concentration of the recycled fragments. The recycle is successes as the concentration is considerable.

 

26.7.2023

Ecvents:The enzymIc linkage was down and the plasmid was transformed in to the competent cell .

 

27.7.2023

Events:Extract and digest the plasmid, the graph of gel under the UV lamp shows two strips which mean the target plasmid was constructed. The E.coli with plasmid was them moved to large volume of LB and cultivated.

 

28.7.2023

Events:Prepared the solution IPTG to inducing the protein expression.

 

29.7.2023

Events:Nickel column purification and nickel column recovery to get the target protein.

 

30.7.2023

Events:Construct SDS-PAGE and do anion chromatography.

 

31.7.2023

Events:EMSA detection of protein and DNA binding.

 

1.8.2023

Events:Detect the cleavage ability of protein.

 

12.9.2023-22.9.2023

Events: Nickel column purification of protein sva/asp, SDS-PAGE and EMSA