Here is the summary of all the experiments iGEM CSMU_Taiwan 2022 designed and conducted this year. Overall, our experiments can be divided into four parts:
Although we only made it to the third session (ELISA) due to the ongoing COVID pandemic in our country, we were still optimistic about the influence of our results and would love to share what we had done with all the iGEMers! For more details, please refer to the bars below or see the Proof of Concept page. As for the connection between our wet lab experiments and dry lab modeling, please see the Design page and Model page.
RPMI(with Glutamine & NaHCO3), FBS 10%, antibiotics (penicillin and streptomycin) 1% Pipet, pipet aid, pipet tip, flask, cell needed (frozen in the liquid Nitrogen)
Cell suspension, trypan blue, a hemocytometer or a Coulter counter, a microscope, phosphate buffer saline (PBS), a centrifuge
(Total cell number in the 4 large squares) x 2 x 10^4 / 4 = cells/ml
Lentivirus, target cells, a 6-well plate, RPMI/IL-2(IL-2 10ng/ml), lenti-GFP(MOI=10), stock solution of the polybrene, an incubator, a centrifuge
A centrifuge, a small centrifuge, target cells, iced phosphate buffer saline (PBS), lysis buffer, protein inhibitor, microcentrifuge tubes (Eppendorf tubes), a pipet, a vortex
The Western blot apparatus, three clean centrifuge tubes, ddH2O, glass plates(large and small), the gel casting stand, sponge, Vaseline, solution A, solution B, solution C, 10% SDS, 10% APS, a 15-well comb, explosion-proof clips, Eppendorf tubes, a small centrifuge, a vortex, a gel holder, a tank, fresh running buffer, reused running buffer, the marker, ice.
Name | running gel(ml) | stecking gel(ml) |
---|---|---|
ddH20 | 6.66 | 4.2 |
Solution A | 5.33 | 0.9 |
Solution B | 4 | X |
Solution C | X | 1.687 |
10% SDS | 0.6 | 0.0675 |
10% APS | 0.08 | 0.0337 |
An iron plate, RO water, alcohol, reused transfer buffer, the black fleece layer, the 'sandwich' cassette, the filter paper, the PVDF membrane (activated), the gel that has finished running, a Styrofoam box, ice, the transfer tank, milk powder, TBST, Eppendorf tubes, a plastic box, a vortex, a shaker, a plastic dropper
This part takes three days to complete.
solution A, solution B, ddH2O, a tweezer, alcohol, the PVDF membrane, the gel imaging machine
Name | dosage |
---|---|
solution A | 500μl |
solution B | 500μl |
ddH2O | 3000μl |
ELISA plate(96 well), the antigen, the coating buffer B, a shaker, TBST, BSA, Anti-Mouse IgG conjugate HRP in blocking buffer, sulfuric acid
Hybridoma cells, RNA extraction reagent, reverse transcriptase, dNTPs, H2O, RNAse inhibitor, primers for PCR, PCR buffer, polymerase, TAE buffer, PCR Clean-Up and Gel Extraction kit
Meyer L, López T, Espinosa R, Arias CF, Vollmers C, DuBois RM (2019) A simplified workflow for monoclonal antibody sequencing. PLoS ONE 14(6): e0218717. https://doi.org/10.1371/journal.pone.0218717