plant DNA extraction CTAB protocol - - (Oct/07/2004 )
I have been tring to extract genomic DNA from sugarcane using CTAB but never succesful. In fact once in awhile I get good quality DNA but very irregularly. I wonder if CTAB can degrade DNA, it obviously ppt. with DNA, is't it. Please any body have any idea what's wrong??
I havn't done this for about 5 years so I am a little rusty on the chemistry, but I think CTAB is in the proceedure to remove polyphenolics from the plant cell walls. Polyphenolics are long chain compounds similar in structure and size to DNA and will precipitate under the same conditions. PVP(polyvinylpyrrilidone) will also help remove these compounds.
I suspect however that you are having problems isolating DNA because of the sugars co-precipitating with the nucleic acids (which are sugars anyway) and sugarcane is bound to have lots of sugar in it. I used this portocol when I was doing plant DNA extractions on Pittosporum, a pretty sugary plant also...www.flmnh.ufl.edu/soltislab/Soltis%20CTAB1.pdf
Hope that helps.
hi, funny you should ask for sugercane. in my search for a high put through extraction protocol i came across this paper.
"An Improved and Rapid Protocol for the Isolation of Polysaccharide- and Polyphenol-Free Sugarcane DNA"
by S. M. ALJANABI , L. FORGET and A. DOOKUN
i am in the process of trying it out on red cherry samples, hope it helps