I'm trying to extract DNA from 70% ethanol fixed cell suspensions.
Firstly, The cells are fixed and stained for cell cycle analysis and collected after flow cytometry. the question is that the cells are hard to be spin down. I can't see the cells on the bottom of the tube after centrifuge. (I am sure that the sorted cells do suspend above the bottom in the liquid)
How to spin the cells down? If can't, how to extract DNA from suspension cell in 2-3ml PBS?
Any suggestions would be appreciated.
How many cells did you collect? You might not see a pellet becuase there are just too little cells?
The other thing to try is to increase you centrifuge speed. The cells are fixed... so there shouldn't be any issues if you overspin a bit.
It is possible to just add a lysis buffer to the 2-3mL PBS... It's probably best to use a qiagen kit since you can concentrate the DNA in the spin columns.
Yes, I am agree with Shen-An Hwang. You can spin the cells down at 500 g for 30 minutes or even more speed. Then can follow the extraction kit guidelines...
Good Luck!
Dear Liu Fang
For best result, you could centrifuge sample in high speed (13.3 rpm for 30 min) and then discard the superficial fluid. After you follow the instructions extraction kit.
Good luck
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