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how to remove brown pigment in Sargassum DNA isolation?

sargassum dna brown pigment dna isolation algae dna isolation

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#1 ajie

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Posted 13 December 2012 - 04:21 AM

Dear all,

I've been doing DNA isolation for "seaweed DNA fingerprinting project". I use brown algae Sargassum DNA as the subject. But the DNA pellet always come with brown color. I suspect the pigment is coprecipitated with the DNA pellet. I have used chloroform incubation and also doing repetitive ethanol washing on the pellet. but it didn't work. Does anyone know how to overcome this problem?

Thank u.

Ajie

#2 hobglobin

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Posted 13 December 2012 - 10:00 AM

I had this sometimes with some insect species, using a salting out protocol and a kit....both had the same result. But it did not influence the work (PCRs only).

One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

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#3 phage434

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Posted 13 December 2012 - 12:24 PM

Apart from aesthetics, what is the problem? You could try adding activated charcoal while it was in suspension, then filtering and precipitating.

#4 azadeh12345

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Posted 14 December 2012 - 07:09 AM

Thank you, How can I add charcoal? could you please suggest me a protocol?
In fact, I have problems with PCR, I guess it may be due to the pigments. Or maybe there is other inhibitors, I am confused

#5 hobglobin

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Posted 14 December 2012 - 09:01 AM

Here they describe how to extract DNA and remove inhibitors in brown algae.
Also in this paper in the attachment, they attribute inhibition to large amounts of polysaccharides.
Attached File  Varelaetal2006Jphyc.pdf   203.27KB   191 downloads

One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

That is....if she posts at all.






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