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DNA purification


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3 replies to this topic

#1 mic

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Posted 19 April 2010 - 02:10 AM

Hi,

I have measured the DNA concentration and the optical density is quite low (~1,6). The DNA is dissolved in water. Is it possible to precipitate it with isopropanol, spin it down and wash it with ethanol? Is there a better way to do that?

#2 Curtis

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Posted 19 April 2010 - 04:50 AM

Hi,

I have measured the DNA concentration and the optical density is quite low (~1,6). The DNA is dissolved in water. Is it possible to precipitate it with isopropanol, spin it down and wash it with ethanol? Is there a better way to do that?


if you have added only 30ul of water in the end of purification step then just top up your sample with 1.5 ml IsoP, incubate at RT for 5 min and centrifuge at 13000 RPM for 10-15 min.

Why do you want to try other methods? this is the easiest and fastest way. IsoP and ethanol are available in any lab.

#3 phage434

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Posted 19 April 2010 - 05:00 AM

Quite low for what? That is a very reasonable concentration of DNA. You can precipitate and wash, but will lose some DNA.

#4 ProteinWork

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Posted 19 April 2010 - 06:42 AM

Hi,

I have measured the DNA concentration and the optical density is quite low (~1,6). The DNA is dissolved in water. Is it possible to precipitate it with isopropanol, spin it down and wash it with ethanol? Is there a better way to do that?

OD 1.6 equals roughly 80ng/ul for double-stranded DNA. That's not low at all. It might not be the highest yield for a miniprep, but definitely within the normal range I've seen. For most applications I know, that concentration is more than enough.




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