In my lab, most scientist had max rcf(about 15000g?) for centrifuging cells when they performed RNA precipitation with Trizol. And cells are about 1*10^5 ~ 1*10^6. How much g force is the least damage to cells?
Hello Kong Yeonghyeon
To extract RNA from cells using TRIzol, if there is a lot of insoluble material after a 5-minute room temperature incubation with TRIzol, the cell lysate is centrifuged at 12000-15000g for 10-15 mins at 4 degree C to remove cellular debris before adding chloroform.
As far as g force is concerned that can cause least damage to cells, you may centrifuge the cells between 200 and 900 RCF, with 900 RCF being more suitable for fixed cells and the lower force values being used for live, unfixed cells. Centrifuging cells at too high of a speed can lead to compact pelleting and damage to the cell membrane, while centrifuging at too low of a speed will allow cells to remain in suspension and be lost when the supernatant is aspirated. I would suggest that you do not exceed 300 RCF for 5-7 mins to pellet the cells without causing damage to the cells.
Best.
The g-force (relative centrifugal force, RCF) required to pellet cells depends on the type and size of the cells. Here are general guidelines for common cell types:
- Bacterial cells: 3,000–10,000 × g for 5–10 minutes
- Yeast cells: 3,000–5,000 × g for 5–10 minutes
- Mammalian cells: 300–500 × g for 5–10 minutes (to avoid damaging the cells)
- Algal cells: 3,000–5,000 × g for 5–10 minutes
For soil bacterial communities, centrifugation at 6,000–10,000 × g for 5–15 minutes is commonly used to pellet the cells efficiently.
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