Team:Shanghai United/Notebook

The Filters

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Notebook

 

Day1.

• Learning the project and related knowledge;
• Ice breaker games;
• Design team name and logo

 

Day2.

• Laboratory safety training;
• Prepare LB culture media, sterilized;
• Cultured the strains containing pET28a-vector plasmid and p15a-vector plasmid respectively;

 

Day3.

• Obtained PPO and P14- tyrR- Ptyr genes through PCR amplification, agarose gel electrophoresis, and gel extraction;
• Extracted the plasmid pET28a-vector and p15a-vector respectively;

 

Day4.

• Enzyme digested the plasmid to obtain the linearized vector;
• Connected the DNA fragment and plasmid-vector and transformed into strain;
• Prepared kanamycin and chloramphenicol solution;

 

Day5.

• Pick bacteria, clone PCR identification;
• Sequence assay;
• Culture the positive clone;

 

Day6.

• Extracted the constructed plasmid;
• Transformed the plasmid into strain;

 

Day7.

• IPTG induced the protein expressing;
• Tyr induced the GFP protein expressing;

 

 

Day8.

• Purified PPO protein;
• SDS-PAGE Gel Electrophoresis;
• Measured the fluorescence intensity of GFP;

 

Day9.

• Polyphenol oxidase (PPO) quantification;
• Tyr induced the PPO protein expressing;

 

Day10.

• Quantified the degradation of p-cresol by PPO；
• Collected the data.

 

 

 

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