Notebook
7/1
Ice breaking for the team
Introduction to the project
Discussion team name and logo design
7/2
PCR to amplify the target gene
Extract pET28(+) plasmid from Escherichia coli DH5-alpha
7/3
Liquid LB medium
Gel electrophoresis and gel extraction
Link vector and target gene by Gibson ligase
Vector transport target gene into competent cell
7/4
Pick bacteria after PCR
Gel electrophoresis test
Extraction of plasmid
Plasmid double digestion
Transport recombinant plasmid into BL21(DE3)
Plate on agar plate
7/5
Pick bacteria and PCR
Gel electrophoresis
Sample sequencing
Cultivate the bacteria
Prepare IPTG for inducing the protein expression
Prepare buffer A and buffer B for protein purification
7/6
Secondary enlargement of bacteria
IPTG induce protein expression
Prepare the gel for SDS-PAGE
7/7
Protein purification
SDS-PAGE
7/8-7/9
Add different concentrations of protein to 96-well plate
Add bacteria E.coli into different holes
Constant temperature shake cultivate 12 hours
Put into the microplate reader to observe the results
7/10-7/11
The second time sterilization test
Add different concentrations of protein to 96-well plate
Add cultured mixed bacteria from water samples from the aquarium into different holes
Constant temperature shake cultivate 12 hours
Put into the microplate reader to observe the results