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Silica magnetic beads can bind nucleic acids controlled by three competing effects: weak electrostatic repulsion forces, dehydration, hydrogen bond formation(Melzak et al., 1996). By the support of guanidine salts, detergents, or proteinase K, the nucleic acids are released from the biological samples, such as cell, tissues, plant and microorganism. Consequently, the released nucleic acids specially bind to silica magnetic beads forming DNA/RNA-beads complexes under the condition of chaotropic salts (e.g. guanidine salts, perchlorate salts and isopropanol etc.). Through magnetic separation, the complexes can be rapidly collected from the biological sample lysis solution. Proteins, other impurities and any salts are washed away by several washing steps with different washing buffers. Finally, the pure DNA/RNA can be easily eluted from the beads by TE(10mM Tris, 1mM EDTA, pH8.0) or ddH2O(pH>7.0). From the basic principle described above, we can get the optimal reagents and operating process by adjusting parameters including pH value, salt concentration, kind of chaotropic or kosmotropic salt, detergent concentration, which involve the lysis buffer, binding buffer, washing buffer and elution buffer.
At present, the chemical constituents in nucleic acids isolation kits differ from different company, but all these kits are mainly divided into two groups, chaotropic salt system(Boom et al., 1990), kosmotropic salts system(Lee et al., 2008), according to the salts used for binding. The chaotropic salts, such as iodized salts, perchlorate, and guanidine salts, which have the ability to disrupt the regular hydrogen bond structures. The high concentration of chaotropic salts let proteins denature, nucleicacidase lose activity, and help silica magnetic beads selectively bind nucleic acids. In contrast, the Kosmotropic salts have much stronger interactions with water and promote hydrophobic interactions, these including sodium sulfate and ammonium sulfate, etc.
The novel method of nucleic acids isolation based on silica beads can be used to extract DNA/RNA with different length from wide varieties of biological samples. For examples, isolation genomic DNA from plant(Zhang et al., 2007), fungi(Rittich et al., 2006), blood(Duarte et al., 2010), forensic tissue(Nagy et al., 2005); and isolation RNA from cells(Lee et al., 2008), plant(Ding et al., 2008); and isolation plasmid DNA from bacterial cells(Chiang et al., 2006); meanwhile certain reagents match silica magnetic beads for DNA/RNA isolation also listed in these referred publications.
Fig. The sketch of application of silica magnetic beads on nucleic acids isolation
References
Boom R., Sol C., Salimans M., Jansen C., Wertheim-van Dillen P., Van der Noordaa J. (1990) Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28:495.
Chiang C.L., Sung C.S., Chen C.Y. (2006) Application of silica-magnetite nanocomposites to the isolation of ultrapure plasmid DNA from bacterial cells. Journal of Magnetism and Magnetic Materials 305:483-490.
Ding L.-W., Sun Q.-Y., Wang Z.-Y., Sun Y.-B., Xu Z.-F. (2008) Using silica particles to isolate total RNA from plant tissues recalcitrant to extraction in guanidine thiocyanate. Analytical Biochemistry 374:426-428.
Duarte G.R.M., Price C.W., Littlewood J.L., Haverstick D.M., Ferrance J.P., Carrilho E., Landers J.P. (2010) Characterization of dynamic solid phase DNA extraction from blood with magnetically controlled silica beads. Analyst 135:531-537.
Lee M., Huh N., Kim J.H. (2008) Isolation of total RNA from Escherichia coli using kosmotropic Hofmeister salts. Analytical Biochemistry 381:160-162. DOI: Doi 10.1016/J.Ab.2008.06.015.
Ligozzi M., Fontana R. (2003) Isolation of total DNA from bacteria and yeast. African Journal of Biotechnology 2:251-253.
Melzak K., Sherwood C., Turner R., Haynes C. (1996) Driving forces for DNA adsorption to silica in perchlorate solutions. Journal of Colloid and Interface Science 181:635-644.
Nagy M., Otremba P., Krüger C., Bergner-Greiner S., Anders P., Henske B., Prinz M., Roewer L. (2005) Optimization and validation of a fully automated silica-coated magnetic beads purification technology in forensics. Forensic Science International 152:13-22.
Rittich B., Spanova A., Horak D., Benes M.J., Klesnilova L., Petrova K., Rybnikar A. (2006) Isolation of microbial DNA by newly designed magnetic particles. Colloids and Surfaces B-Biointerfaces 52:143-148. DOI: DOI 10.1016/j.colsurfb.2006.04.012.
Zhang Z.C., Cui Y., Wan Q.H. (2007) Surface modification of magnetic silica microspheres and its application to the isolation of plant genomic nucleic acids. Chinese Journal of Analytical Chemistry 35:31-36.