Event: Ice breaking games and team discussions
Agenda: Self-introduction, introduction of the program and rules, brainstorm of team name, logo, slogan, and the team uniform.
logo and name:
Event: Lab day 1
Agenda: Laboratory safety training, use of pipette gun, allocation of reagents, manipulation and use of precision instruments, allocation of LB medium, PCR manipulation and use, shaken plasmid, agarose gel electrophoresis.
Purpose: Amplified NS3-Hel and NS3-ZIKA genes together with pET28a and constructed the plasmids by instrumentation.
Event: Lab day 2
Agenda: Plasmid extraction and concentration determination, DNA tool enzyme digestion, ligand, transformation of DH5α cells, plate coating screening.
Purpose: After enzyme digestion, the target fragment of NS3-Hel/NS3-ZIKA was obtained by PCR, and the recombinant plasmid was obtained by enzyme-linking.
Event: Lab day 3-4
Agenda: The transformers were identified by PCR, the monoclonal seed solution was selected, and the plasmids pET28a-NS3hel and ET28a-NS3-ZIKA were extracted and transformed into BL21(DE3).
Purpose: Prove that the plasmid solution contain NS3-Hel and NS3-ZIKA by comparing the magnitude of the stripes, and transfer the DNA recombinant plasmid into BL21(DE3).
Event: Lab day 5
Agenda: Isolation of single bacteria, PCR identification of inverters, agarose gel electrophoresis verification of target bands, isolation of monoclonal culture seed solution, preparation of protein purification buffer.
Purpose: The purpose of running a gel and performing PCR validation after amplification is to verify the presence of target bands for NS3-Hel and NS3-ZIKA, and to inoculate and validate the correct colonies, and to perform pre-culture of the bacterial suspension for subsequent experiments.
Event: Lab day 6-7
Agenda: Expanded culture seed solution, IPTG induced expression, SDS-PAGE protein glue preparation.
Purpose: To trigger the expression of the target gene or protein in the transformed cells.
Event: Lab day 8-9
Agenda: Ultrasonic crushing, nickel column affinity purified protein, SDS-PAGE protein electrophoresis.
Purpose: Filter or purify the target protein from other protein.
Event: Lab day 10-11
Agenda: SDS-PAGE protein electrophoresis, protein glue staining and decolorization
Purpose: Validation protein expression.
Event: Lab day 12-13
Agenda: Functional test - enzyme marker test
Purpose: Test the function.
Event: Lab day 15
Agenda: Functional test II - enzyme marker test
Purpose: Duplicate the function test by adding controls.
Event: Lab day 16
Agenda: WIKI&PARTS key points interpretation and task division, organization of writing WIKI, organization of uploading PARTS
