I mean how is it going to precipitate the polysaccharides? Does it precipitate +ve or -vely charged proteins? What about polysaccharides?
Usually during genomic DNA isolation from plants we use CTAB that selectively precipitates nucleic acids. RNA and DNA are insoluble complex with CTAB at low salt strength (eg. 0.4 M NaCl) but the CTAB-Nucleic acid complex becomes soluble at high salt concentration (0.7 M NaCl). But many polysaccharides are insoluble at this salt range are not solubilized thus remain precipitated.
Refer: M.G.Murray and W.F.Thompson 1980.
Good Luck
Neelam
I am little bit sceptical about the capacity of CTAB to solubilize the plant cell wall. It is our experience that the cell wall should always be mechanically discrupted prior to nucleic acids extraction. After the disruption, CTAB certainly can separate polysachararides from DNA/RNA. Importantly, CTAB is the probably the only compound that can separate, at least, partially nucleic acids from polyphenols. The polyphenolic compounds may severly inhibit downstream DNA/RNA reactions.
The CTAB high salt buffer and elevated incubation temperature /60C/ work best. I do nor recomment precipitation of RNA/DNA CTAB complexes at low ionic strenght since the complexes are very difficult to dissolve.
Thank u sir....................m trying to extract genomic DNA from Amaranthus plant leaf,but not able to get it...........m trying to fix the problem. m using PVPP 40(range) is it ok?
Dear Bhagya,
I have no experience with polyvinylpyrolidone-assisted DNA extraction. However, I am sending two our CTAB protocols that worked fine with most plant DNA extractions.Ales
Dear sir Ales Kovarik
Thank u very much i will try this method.
Finally I got the genomic DNA .......thanks for everyone who helped me through protocols and answers.
Ales, are you able to quantify the concentration of CTAB extracted DNA using a dye such as SYBR Green?
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