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Lab notebooks

2024/07/15-2024/07/19

  1. Conducting laboratory safety training and safety assessments

  2. Conduct basic experimental operations to cultivate experimental thinking and abilities

  3. Simultaneously complete the experiment of blue and white class screening(Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)

2023/07/22-2023/07/26

Construction of plasmid pCDFDuet-1-GFP and PETDuet-1-nativeP-alsR

  1. Use PCR to amplify a bidirectional promoter fragment on the genome of Escherichia coli bl21.
  2. Using pCDFDuet-1-GFP DPE as a template, amplify the linearized vector pCDFDuet-1-GFP by PCR.
  3. Obtaining plasmid pCDFDuet-1-Palsl-sfGFP through Gibson assembly.
  4. Use PCR to amplify an alsR fragment on the genome of Escherichia coli bl21.
  5. Using pETDuet-1 as a template, amplify the linearized vector pETDuet-1 by PCR.
  6. Obtaining plasmid pETDuet-1-nativeP-alsR through Gibson assembly.
  7. Double plasmid transformation: pCDFDuet-1-Palsl-sfGFP and pETDuet-1-nativeP-alsR. (Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)

2024/07/29-2024/08/02

Construction of plasmid pCDFDuet-1-alsR-P-sfGFP1 and fluorescence measurement of bacterial cultures

  1. Dual-plasmid Biosensor Performance Verification and Fluorescence measurement of bacterial cultures.
  2. PCR amplification of the alsR fragment on plasmid pETDuet-1-p-alsR.
  3. Using pCDFDuet-1-palsl-sfGFP as a template, PCR amplification was performed on the linearized vector pCDFDuet-1-p-sfGFP
  4. Obtaining plasmid pCDFDuet-1-alsR-P-sfGFP through Gibson assembly.
  5. Construction results and performance verification of pCDFDuet-1-alsR-P-SfGFP1.
  6. Fluorescence value determination of single plasmid system culture. (Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)

2024/08/05-2024/08/09

  1. Conducting laboratory safety training and safety assessments
  2. Conduct basic experimental operations to cultivate experimental thinking and abilities
  3. Simultaneously complete the experiment of blue and white class screening (Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)

2024/08/12-2024/08/16

  1. PCR amplification of the alsR fragment on plasmid pETDuet-1-p-alsR.
  2. Using pCDFDuet-1-palsl-sfGFP as a template, PCR amplification was performed on the linearized vector pCDFDuet-1-p-sfGFP
  3. Performing Error-Prone PCR on the Target alsR Fragment.
  4. Obtaining mutated plasmid pCDFDuet-1-alsR-PsfGFP through Gibson assembly.
  5. Performance validation and fluorescence value determination of mutant plasmid system culture (Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)

2023/08/19-2023/08/23

  1. PCR amplification of the CtRpiB fragment on bl21 genome.
  2. Using pET-28a(+) plasmid as a template, PCR amplification was performed on the linearized vector pET-28a(+).
  3. Obtaining plasmid pET-28a(+)-CtRpiB through Gibson assembly.
  4. Construction of dual plasmid pET-28a (+) - CtRepiB and mutant plasmid.
  5. Induction and performance validation of dual plasmid biosensors and fluorescence measurement of bacterial cultures. (Experimenter: Lin Li、Yi Cao、Yuting Lu、Chenxi Lu、Fang yuan、Ying Zhang)