Comparative characteristics of sorption methods of bacterial and fungal DNA isolation from pollen
Abstract
DNA and RNA isolation methods, based on the binding of nucleic acids with sorbent media are widely used at present time. The efficiency of bacterial, fungal and plant DNA isolation method from pollen has not been studied. We have studied the effectiveness of commercial kits – «Proba-GS»; «Nukleosorb, Komplektacia "C''» and «Quick-gDNA ™ MiniPrep», as well as of the traditional method of DNA extraction with organic solutions, on efficiency of bacterial and fungal DNA isolation from pollen collected by bees. It was shown that the methods based on the adsorption of nucleic acids completely eliminate contamination by RNA. The highest concentration of DNA was obtained using «Nukleosorb, Komplektacia "C''» kit. Analysis of the relative gene expression in bacteria, plants and fungi revealed no significant differences in the isolation efficiency by different methods.
Downloads
References
Barnett R., Larson G.A., Methods Mol. 6. Selemenev V.F., Rudakov O.B., Biol., 2012, Vol. 840, pp 13-19. Slavinskaja G.V., Drozdeva N.V., M., DeLi
2.
Cady N.C., Stelick S., Batt C.A., Biosens print, 2008, 246 p. Bioelectron., 2003, Vol. 19, pp. 59-66. 7. Karpov S.I., Matveeva M.V., Selemenev
3.
Melzaka K.A., Sherwooda C.S., Turnerb V.F., Russian Journal of Physical Chemistry A. R.F.B., Haynesc C.A., J. of Colloid and 2001, Vol. 75, No 2, pp. 266-271. Interface Science, 1996, Vol. 181, No 2, 8. González G., Hinojo M.J., Mateo R., pp 635-644. Medina A. et al., Int. J. Food Microbiol, 2005,
4. Poeckh T., Lopez S., Fuller A.O., Solomon Vol. 105, pp. 1-9.
M.J. et al. // Anal Biochem. 2008. Vol. 373, pp. 9. Kacániová M., Fikselová M., Ann. Agric 253-262. Environ Med., 2007, Vol. 14, pp. 229-232.
5. Kendall D., Lye G.J., Levy M.S., Biotechnol Bioeng., 2002, Vol. 79, pp. 816-822
10. Hebert P.D., Ratnasingham S., deWaard J.R., Proc. Biol. Sci.., 2003, Vol. 270, pp. 9699.
11. Schoch C.L., Seifert K.A., Huhndorf S., Robert V. et al., Proc. Natl. Acad. Sci., 2012, Vol. 109, pp. 6241-6246.
12.
Dong W., Cheng T., Li C., Xu C. et al., Mol. Ecol. Resour., 2014, Vol. 14, No 2, pp. 336-343.
13.
Sameer A., Barghouthi A., Indian J. Microbiol., 2011, Vol. 51, pp. 430-444.
