Team:Concordia/Safety/SOP/Nanoshell
Nanoparticle Attachment: Nanoshell Method for Yeast
*The safety information in the table above is based on the SDS for each chemical.
1. Sigma-Aldrich Corporation. SDS: Trisodium Citrate Dihydrate.
Can be used to attach gold nanoparticles synthesized from Martin method, garlic synthesis method or purchased from Cedarlane. Attachment to yeast only.
Cells: Saccharomyces cerevisiae Risk group: Level 1 (https://2014.igem.org/Safety/Risk_Group_Guide)
Protocol for attaching 5nm gold nanoparticles to yeast cells:
1. Measured 0D600 of cells = 10.4
2. Transferred 2mL of cells to a 2mL Eppendorf tube and centrifuge at 5000rpm for 20 minutes to collect cells.
3. Decant supernatant, resuspend in 1mL of PBS
4. Centrifuge at 5000rpm for 20 minutes
5. Decant supernatant, resuspend in 1mL of PBS
6. Set up 5 tubes according to the following table:
| % NP | 0% | 20% | 40% | 60% | 80% |
|---|---|---|---|---|---|
| Vol yeast | 100 µL | 100 µL | 100 µL | 100 µL | 100 µL |
| Vol NP | 0 | 400 µL | 800 µL | 1.2 mL | 1.6 mL |
| Vol PBS | 1.9 mL | 1.5 mL | 1.1 mL | 700 µL | 300 µL |
*Vol yeast corresponds to the volume of cell solution taken from the tube containing PBS/yeast (Step 5)
7. Incubate at room temperature for half an hour
8. Centrifuge at 4000rpm for 15 minutes
9. Resuspend in 1mL of 1X TE buffer
Source: Jiang, N., Yang, X-Y., Ying, G-L., Shen, L., Liu, J., Geng, W., Dai, L-J., Liu, S-Y., Cao, J., Tian, G., Sun, T-L., Li, S-P., and Su, B-L. (2015) “Self-repairing” nanoshell for cell protection. Chem Sci., 6:486-491. Retrieved from http://pubs.rsc.org/en/Content/ArticleLanding/2015/SC/c4sc02638a#!divAbstract
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