Extracting DNA from FACS sorted cellsDNA after FACS
Posted 20 September 2011 - 09:07 PM
I am sorting a rare population of cells by cytometry... I am getting the sorted cells in a volume of 5 ml... I have tried this a couple of times to make a cellular pellet from this 5 ml... Unfortunately, I am loosing a lot of cells in the supernatant... Thus my yield of DNA is very small... Can anyone please tell me, how I could get the cell pellet without losing much cells after cytometry... I would be extracting DNA from this pellet...
I will appreciate any help in this regards...
Posted 20 September 2011 - 10:13 PM
Posted 21 September 2011 - 08:07 AM
On the other hand, as you are going to lyse your cells to extract the DNA, I assume viability is not an issue, so you could always use harsher ways. You can centrifuge them in a benchtop micro-centrifuge. Either split the 5ml in 5 x1.5ml tubes, or spin 1ml at a time. 5min at max speed should definitely pellet the cells. Remove the supernatant, and resuspend in lysis buffer. I do that to prepare lysates for protein analysis and so far has never failed me.
Posted 28 November 2011 - 08:21 PM