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SC2-VLP concentration using PEG precipitation
Last updated date: Dec 1, 2022 Views: 1559 Forks: 0
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SC2-VLP concentration using PEG precipitation
- Prepare PEG precipitation solution using 50% w/v polyethylene glycol 8000, 2.2% NaCl.
- For each 100mL of VLP containing supernatant, add 13.63mL of precipitation solution. Final concentration will be 6% PEG. Mix thoroughly and cool for 2 hours at 4C. Incubations longer than 3hrs will result in debris and difficult to resuspend pellet.
- Centrifuge at 2000g for 20 minutes. Remove the majority of supernatant and spin again at 2000g for 1 minute and remove the remaining supernatant.
- Resuspend pellet in 1/100th volume of PBS. Pellet may require up to 10 minutes to resuspend.
- Can proceed with infection on the same day or freeze and store at -80C.
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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
- Syed, A, Ott, M and Doudna, J(2022). SC2-VLP concentration using PEG precipitation. Bio-protocol Preprint. bio-protocol.org/prep2072.
- Syed, A. M., Taha, T. Y., Tabata, T., Chen, I. P., Ciling, A., Khalid, M. M., Sreekumar, B., Chen, P., Hayashi, J. M., Soczek, K. M., Ott, M. and Doudna, J. A.(2021). Rapid assessment of SARS-CoV-2 evolved variants using virus-like particles. Science. DOI: 10.1126/science.abl6184
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