Inhibition of random amplified polymorphic DNAs (RAPDs) by plant polysaccharides
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A survey of the inhibition of the amplification of spinach DNA by various plant polysaccharides revealed that neutral polysaccharides (arabinogalactan, dextran, gum guar, gum locust bean, inulin, mannan, and starch) were not inhibitory. In contrast, the acidic polysaccharides (carrageenan, dextran sulfate, gum ghatti, gum karaya, pectin, and xylan)were inhibitory. In the process of preparing random amplified polymorphic DNAs (RAPDs), the loss of large DNA bands appears to be an indicator that the fingerprint pattern has been affected by polysaccharides. The addition of various concentrations of Tween 20, DMSO, or PEG 400 to the PCR reaction mixture resulted in partial restoration of amplification of RAPDs for the acidic polysaccharides. The most effective way to eliminate the effects of polysaccharide inhibition was by diluting the DNA extracts, and thereby diluting the polysaccharide inhibitors.
Key WordsRAPDs PCR polysaccharides inhibition plants buffer enhancers
random amplified polymorphic DNA
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- Caetano-Anolles, G., B.J. Bassam and P.M. Gresshoff. 1994. Multiple arbitrary amplicon profiling using short oligonucleotide primers, pp. 29–45 inPCR Technology: Current Innovations. (eds. H.G. Griffin, A.M. Griffin). CRC Press, Boca Raton.Google Scholar
- Demeke, T., and R.P. Adams. 1992. The effect of plant polysaccharides and buffer additives of PCR. Bio Techniques 12:332–334.Google Scholar
- Demeke, T., and R.P. Adams. 1994. The use of PCR-RAPD analysis in plant taxonomy and evolution, pp. 179–192 inPCR Technology: Current Innovations. (eds. H.G. Griffin, A.M. Griffin). CRC Press, Boca Raton.Google Scholar
- Do, N., and R.P. Adams. 1991. A simple technique for removing plant polysaccharide contaminants from DNA. Bio Techniques 10:162–166.Google Scholar
- Doyle, J.J., and J.L. Doyle. 1987. A rapid DNA isolation procedure for small quantities of fresh quantities of fresh leaf tissue. Phytochem. Bull. 19:11–15.Google Scholar
- Gelfand, D.H. 1989. Taq DNA Polymerase, p. 17–22 inPCR Technology. (ed. H.A. Erlich). Stockton Press, New York.Google Scholar
- Pomp, D., and J.F. Medrano. 1991. Organic solvents as facilitators of polymerase chain reaction. Bio Techniques 10:58–59Google Scholar
- Smith, F., and R. Montgomery. 1959. The chemistry of plant gums and mucilages and some related polysaccharides. Reinhold Publishing, New York.Google Scholar