While isolating dna using ctab method, i obtained a gelly ppt (of about 40ul) before i added TE buffer, when the sample was run on agarose gel, i obtained a fairly low conc. of DNA with no smear. can someone explain me the reason behind this error.
Yes, polysacharide and polyphenols contaminants are probably the reason of your jelly-like DNA sample.
-You may try to increase the concentration of CTAB in your extration buffer up to 3% that might help to remove some poylsacharides.
-Also addition of polyvinilidene pyrolidone (PVP) into a standard CTAB extraction buffer helps. We use 2%PVP+1%CTAB buffer to extract DNA from difficult woody tissues.
Finally , the starting of soure material should be increase the yield. Volume of the CTAB lysis buffer can be easily scaled up to as much as 20 ml.
I agree with Ales. To eliminate the polysaccharide and polyphenols you can use phenol/chloroform/isoamyl alcohol (25:24:1) and then chloroform/octanol to eliminate the phenol.
Yes, I agree. Therefore, I always use the Plant DNeasy Kit (Qiagen).
I had never problems (also not with difficult material: Digitalis & Mammillaria).
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