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isolating DNA from cells in suspension


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#1 jamestoon1

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Posted 13 January 2013 - 02:52 AM

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?

#2 leelee

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Posted 13 January 2013 - 05:39 AM

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

#3 jamestoon1

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Posted 13 January 2013 - 06:31 AM

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?

#4 leelee

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Posted 13 January 2013 - 06:40 AM

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

#5 Pangea

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Posted 13 January 2013 - 07:51 AM

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

#6 jamestoon1

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Posted 14 January 2013 - 09:41 PM

Thanks for the feedback!




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