Date: 7th August,2023

We designed team nick-name, logo, and team uniform together. In the afternoon, we also participated a laboratory safety training session.

Date:8th August, 2023

We first extracted the Gox1 gene by PCR and configured the LB medium. Afterwards, we used electrophoresis to examine the final product, but the results showed that the PCR was unsuccessful. So we did the former two steps again and finally succeeded. In the end, we prepared kanamycin sulfate for the next day and poured the solid LB into the plate. Experimental operations are shared equally within the group, with everyone participating in each item.

Date:9th August,2023

Under the instruction of Mr Chen, we recycled the electrophoresis gel we made yesterday whichcontains Goxl gene anddid a PCR again in the morning. Then, we did an electrophoresis on the PCR results. After that, we inoculated and extractedET-28a bacterial fluids.In the afternoon, we recycled the gel and did an enzymatic digestion and ligation of the gene andplasmid. Finally, we put it into the shaker for 16 hours. Experimental operations are shared equally within the group, with everyone participating in each item.

Date: 10th August, 2023

In the morning, we ran an electrophoresis and did gel recovery of the enzymatically cleaved Gox1 gene and extracted the pET28a plasmid from the bacterial fluid. We heated and transformed it into DH5α E.coli and finally cultured the bacteria. Experimental operations are shared equally within the group, with everyone participating in each item.

Date:11th August,2023

In the morning, we picked monoclones of E.coli which we incubated overnight and we performed PCR to identify the result. In the afternoon, we did electrophoresis for the product of PCR. But we did not get a satisfying result, so we have to repeat it tomorrow.Experimental operations are shared equally within the group, with everyone participating in each item.

Date:12th August,2023.

In the morning, we performed PCR on the BL21(DE3) E.coli and amplified E.coli which contained pET28a-Gox1 plasmid. In the afternoon, Mr. Chen taught us how to use Snap Gene and helped us to resuscitate 4T1 breast cancer cells. Then, we used IPTG to induce protein expression. Experimental operations are shared equally within the group, with everyone participating in each item.

Date: 13th August, 2023

In the morning, we centrifuged and used ultrasound cell crusher to crush BL21(DE3) E.coli induced overnight. In the afternoon, we used nickel columns to purify the proteins we got and configured the SDS-PAGE protein gel. Experimental operations are shared equally within the group, with everyone participating in each item.

Date:14th August,2023

In the morning, we first examined glucose oxidase activity and did SDS-PAGE on the protein we purified yesterday.

In the afternoon, we used coomassie brilliant blue staining to test protein quantification and decolorized it.

In the late afternoon,with the help of Mr. Chen, we examined CCK-8 of 4T1cells.Experimental operations are shared equally within the group, with everyone participating in each item.

Date:15th August ,2023

In the morning, all of our dry and wet team members gathered together to set photos and decided the final team lead is Sishuo Ye.In the afternoon, Qirui Du and Shengfan Jin went to the dry team to help teach children about cancer in a neighborhood.

The other members of the wet lab team stayed in the lab to run SDS-PAGE again and tested the reduced glutathione content(GSH activity detection). Experimental operations are shared equally within the group, with everyone participating in each item.

Date:16th August,2023

--One Day off

Date:17th August,2023

In the morning,all of us did preparations for the presentation and decided division of labor for the wiki. In the afternoon, we had a training lesson about how to do a presentation and decided on each member’s part in the presentation. Also some members went to the laboratory for final check and clean.

Date: 18th August, 2023

In the morning, we did the internal rehearsal presentationand continue our wiki documentation including result data analysis.