5 replies to this topic

[jamestoon1]

When isolating DNA from cells in suspension, most if not all the protocols I have come across recommend centrifugating the suspension at the very beginning to obtain a cell pellet, followed by resuspension of the pellet in either water or PBS or other buffers.

May I know what is the purpose of obtaining this cell pellet?

Can we just vortex the suspension and use it directly without pelleting the cells?

[leelee]

It would be so that you can concentrate your cells into a much smaller volume to facilitate the DNA extraction.

[jamestoon1]

leelee, on 13 January 2013 - 05:39 AM, said:

It would be so that you can concentrate your cells into a much smaller volume to facilitate the DNA extraction.

Oh..so if the protocol recommends us to resuspend the cell pellet into 200ul of buffer, and I have an initial cell suspension volume of 200ul, does it mean that I don't have to perform the centrifugation and resuspension?

[leelee]

You don't really want to be doing an extraction in your cell culture media, so resuspend in buffer like the protocol states.

[Pangea]

Getting rid of unnecessary molecules in media. And handling with less solution (concentrating).

[jamestoon1]

Thanks for the feedback!