4 replies to this topic

[ajie]

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

[hobglobin]

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).

[phage434]

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

[azadeh12345]

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

[hobglobin]

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.
 Varelaetal2006Jphyc.pdf   203.27K   93 downloads