Applied Microbiology and Biotechnology

, Volume 73, Issue 3, pp 495–504 | Cite as

Magnetic particles for the separation and purification of nucleic acids

  • Sonja BerensmeierEmail author


Nucleic acid separation is an increasingly important tool for molecular biology. Before modern technologies could be used, nucleic acid separation had been a time- and work-consuming process based on several extraction and centrifugation steps, often limited by small yields and low purities of the separation products, and not suited for automation and up-scaling. During the last few years, specifically functionalised magnetic particles were developed. Together with an appropriate buffer system, they allow for the quick and efficient purification directly after their extraction from crude cell extracts. Centrifugation steps were avoided. In addition, the new approach provided for an easy automation of the entire process and the isolation of nucleic acids from larger sample volumes. This review describes traditional methods and methods based on magnetic particles for nucleic acid purification. The synthesis of a variety of magnetic particles is presented in more detail. Various suppliers of magnetic particles for nucleic acid separation as well as suppliers offering particle-based kits for a variety of different sample materials are listed. Furthermore, commercially available manual magnetic separators and automated systems for magnetic particle handling and liquid handling are mentioned.


Magnetic particles Nucleic acid Separators Automation 


  1. Akutsu J-I, Tojo Y, Okochi M, Yohda M, Segawa O, Obata K, Tajima H (2004) Development of an integrated automation system with a magnetic bead-mediated nucleic acid purification device for genetic analysis and gene manipulation. Biotechnol Bioeng 86:667–671CrossRefGoogle Scholar
  2. Alderton RP, Eccleston LM, Howe RP, Read CA, Reeve MA, Beck S (1992) Magnetic bead purification of M13 DNA sequencing templates. Anal Biochem 201:166–169CrossRefGoogle Scholar
  3. Bartl K, Wenzig P, Kleiber J (1998) Simple and broadly applicable sample preparation by use of magnetic glass particles. Clin Chem Labor Med 36:557–559CrossRefGoogle Scholar
  4. Birnboim HC (1983) A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol 100:243–255Google Scholar
  5. Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acid Res 7:1513-1523Google Scholar
  6. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503Google Scholar
  7. Boom R, Sol C, Beld M, Weel J, Goudsmit J, Wertheim-van Dillen P (1999) Improved silica-guanidinium thiocyanate DNA isolation procedure based on selective binding of bovine alpha-casein to silica particles. J Clin Microbiol 37:615–619Google Scholar
  8. Bowtell DD (1987) Rapid isolation of eukaryotic DNA. Anal Biochem 162:463–465CrossRefGoogle Scholar
  9. Breadmore MC, Wolfe KA, Arcibal IG, Leung WK, Dickson D, Giordano BC, Power ME, Ferrance JP, Feldman SH, Norris PM, Landers JP (2003) Microchip-based purification of DNA from biological samples. Anal Chem 75:1880–1886CrossRefGoogle Scholar
  10. Bruce IJ, Sen T (2005) Surface modification of magnetic nanoparticles with alkoxy-silanes and their application in magnetic bioseparations. Langmuir 21:7029–7035CrossRefGoogle Scholar
  11. Bruce IJ, Taylor J, Todd M, Davies MJ, Borioni E, Sangregorio C, Sen T (2004) Synthesis, characterisation and application of silica-magnetite nanocomposites. J Magn Magn Mater 284:145–160CrossRefGoogle Scholar
  12. Chiang C-L, Sung C-S, Wu T-F, Chen C-Y, Hsu C-Y (2005) Application of superparamagnetic nanoparticles in purification of plasmid DNA from bacterial cells. J Chromatogr B 822:54–60CrossRefGoogle Scholar
  13. Davies MJ, Taylor JI, Sachsinger N, Bruce IJ (1998) Isolation of plasmid DNA using magnetite as a solid-phase adsorbent. Anal Biochem 262:92–94CrossRefGoogle Scholar
  14. Demeke T, Adams RP (1992) The effects of plant polysaccharides and buffer additives on PCR. Biotechniques 12:332–334Google Scholar
  15. Elaissari A, Rodrigue M, Meunier F, Herve C (2001) Hydrophilic magnetic latex for nucleic acid extraction, purification and concentration. J Magn Magn Mater 225:127–133CrossRefGoogle Scholar
  16. Elaissari A, Ganachaud F, Pichot C (2003) Biorelevant latexes and microgels for the interaction with nucleic acids. Top Curr Chem 227:169–193CrossRefGoogle Scholar
  17. Endres HN, Johnson JA, Ross CA, Welp JK, Etzel MR (2003) Evaluation of an ion-exchange membrane for the purification of plasmid DNA. Biotechnol Appl Biochem 37:259–266CrossRefGoogle Scholar
  18. Fangan BM, Dahlberg OJ, Deggerdal AH, Bosnes M, Larsen F (1999) Automated system for purification of dye-terminator sequencing products eliminates up-stream purification of templates. Biotechniques 26:980–983Google Scholar
  19. Ferreira GNM, Cabral JMS, Prazeres DMF (2000) Studies on the batch adsorption of plasmid DNA onto anion-exchange chromatographic supports. Biotechnol Prog 16:416–424CrossRefGoogle Scholar
  20. Franzreb M, Siemann-Herzberg M, Hobley TJ, Thomas ORT (2006) Protein purification using magnetic adsorbent particles. Appl Microbiol Biotechnol 70:505–516CrossRefGoogle Scholar
  21. Grüttner C, Rudershausen S, Teller J (2001) Improved properties of magnetic particles by combination of different polymer materials as particle matrix. J Magn Magn Mater 225:1–7CrossRefGoogle Scholar
  22. Hawkins TL, O’Connor-Morin T, Roy A, Santillan C (1994) DNA purification and isolation using a solid-phase. Nucleic Acids Res 22:4543–4544Google Scholar
  23. Hei AL, Cai JP (2005) Development of a method for concentrating and purifying SARS coronavirus RNA by a magnetic bead capture system. DNA and Cell Biol 24:479–484CrossRefGoogle Scholar
  24. Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 26:365–369CrossRefGoogle Scholar
  25. Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197CrossRefGoogle Scholar
  26. Horak D, Rittich B, Spanova A, Benes MJ (2005) Magnetic microparticulate carriers with immobilized selective ligands in DNA diagnostics. Polymer 46:1245–1255CrossRefGoogle Scholar
  27. Hourfar MK, Michelsen U, Schmidt M, Berger A, Seifried E, Roth WK (2005) High-throughput purification of viral RNA based on novel aqueous chemistry for nucleic acid isolation. Clinic Chem 51:1217–1222CrossRefGoogle Scholar
  28. Hourfar MK, Schmidt M, Seifried E, Roth WK (2005) Evaluation of an automated high-volume extraction method for viral nucleic acids in comparison to a manual procedure with preceding enrichment. Vox Sang 89:71–76CrossRefGoogle Scholar
  29. Jacobsen N, Nielsen PS, Jeffares DC, Eriksen J, Ohlsson H, Arctander P, Kauppinen S (2004) Direct isolation of poly(A)(+) RNA from 4 M guanidine thiocyanate-lysed cell extracts using locked nucleic acid-oligo(T) capture. Nucleic Acids Res 32:e64CrossRefGoogle Scholar
  30. Levison PR, Badger SE, Dennis J, Hathi P, Davies MJ, Bruce IJ, Schimkat D (1998) Recent developments of magnetic beads for use in nucleic acid purification. J Chromatogr A 816:107–111CrossRefGoogle Scholar
  31. Melzak KA, Sherwood CS, Turner RFB, Haynes CA (1996) Driving forces for DNA adsorption to silica in perchlorate solutions. J Colloid Interface Sci 181:635–644CrossRefGoogle Scholar
  32. Muir P, Nicholson F, Jhetman M, Neogi S, Banatvala JE (1993) Rapid diagnosis of enterovirus infection by magnetic bead extraction and polymerase chain-reaction detection of enterovirus RNA in clinical specimes. J Clin Microbiol 31:31–38Google Scholar
  33. Müller-Schulte D, Schmitz-Rode T, Born P (2005) Ultra-fast synthesis of magnetic and luminescent silica beads for versatile bíoanalytical applications. J Magn Magn Mater 293:135–143CrossRefGoogle Scholar
  34. Obata K, Segawa O, Yakabe M, Ishida Y, Kuroita T, Ikeda K, Kawakami B, Kawamura Y, Yohda M, Matsunaga T, Tajima H (2001) Development of a novel method for operating magnetic particles, Magtration Technology, and its use for automating nucleic acid purification. J Biosci Bioeng 91:500–503CrossRefGoogle Scholar
  35. Olsvik Ø, Popovic T, Skjerve E, Cudjoe KS, Hornes E, Ugelstad J, Uhlén M (1994) Magnetic separation techniques in diagnostic microbiology. Clin Microbiol Rev 7:43–54Google Scholar
  36. Oster J, Parker J, Brassard LÀ (2001) Polyvinyl-alcohol-based magnetic beads for rapid and efficient separation of specific or unspecific nucleic acid sequences. J Magn Magn Mater 225:145–150CrossRefGoogle Scholar
  37. Pichl L, Heitmann A, Herzog P, Oster J, Smets H, Schottstedt V (2005) Magnetic bead technology in viral RNA and DNA extraction from plasma minipools. Transfusion 45:1106–1110CrossRefGoogle Scholar
  38. Prodelalova J, Rittich B, Spanova A, Petrova K, Benes MJ (2004) Isolation of genomic DNA using magnetic cobalt ferrite and silica particles. J Chromatogr A 1056:43–48CrossRefGoogle Scholar
  39. Rolfs A, Weber I (1994) Fully-automated, nonradioactive solid-phase sequencing of genomic DNA obtained from PCR. Biotechniques 17:782–787Google Scholar
  40. Rutjes SA, Italiaander R, van den Berg HHJL, Lodder WJ, de Roda Husman AM (2005) Isolation and detection of enterovirus RNA from large-volume water samples by using the nucliSens miniMAG System and real-time nucleic acid sequence-based amplification. Appl Environ Microbiol 71:3734–3740CrossRefGoogle Scholar
  41. Safarik I, Safarikova M (1999) Use of magnetic techniques for the isolation of cells. J Chromatogr B 722:33–53Google Scholar
  42. Safarik I, Ptackova L, Safarikova M (2001) Large-scale separation of magnetic bioaffinity adsorbents. Biotechnol Lett 23:1953–1956CrossRefGoogle Scholar
  43. Satokari RM, Kataja K, Soderlund H (2005) Multiplexed quantification of bacterial 16S rRNA by solution hybridization with oligonucleotide probes and affinity capture. Microb Ecol 50:120–127CrossRefGoogle Scholar
  44. Spanova A, Horak D, Soudkova E, Rittich B (2004) Magnetic hydrophilic methacrylate-based polymer microspheres designed for polymerase chain reactions applications. J Chromatogr B 800:27–32CrossRefGoogle Scholar
  45. Stadler J, Lemmens R, Nyhammar T (2004) Plasmid DNA purification. J Gene Med 6:S54–S66CrossRefGoogle Scholar
  46. Tan W, Wang K, He X, Zhao XJ, Drake T, Wang L, Bagwe RP (2004) Bionanotechnology based on silica nanoparticles. Med Res Rev 24:621–638CrossRefGoogle Scholar
  47. Teeters MA, Conrardy SE, Thomas BL, Root TW, Lightfoot EN (2003) Adsorptive membrane chromatography for purification of plasmid DNA. J Chromatogr A 989:165–173CrossRefGoogle Scholar
  48. Tian H, Huhmer AFR, Landers JP (2000) Evaluation of silica resins for direct and efficient extraction of DNA from complex biological matrices in a miniaturized format. Anal Biochem 283:175–191CrossRefGoogle Scholar
  49. Ugelstad J, Berge A, Ellingsen T, Schmid R, Nilsen T-N, Sienstad P, Mørk PC, Hornes E, Olsvik Ø (1992) Preparation and application of new monosized polymer particles. Prog Polym Sci 17:87–161CrossRefGoogle Scholar
  50. Ugelstad J, Mork PC, Schmid R, Ellingsen T, Berge A (1993) Preparation and biochemical and biomedical applications of new monosized polymer particles. Polym Intern 30:157–168Google Scholar
  51. Veyret R, Delair T, Pichot C, Elaissari A (2005) Amino-containing magnetic nanoemulsions: elaboration and nucleic acid extraction. J Magn Magn Mater 295:155–163CrossRefGoogle Scholar
  52. Vogelstein B, Gillespie D (1979) Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci USA. 76:615–619CrossRefGoogle Scholar
  53. Vuosku J, Jaakola L, Jokipii S, Karppinen K, Kamarainen T, Pelkonen VP, Jokela A, Sarjala T, Hohtola A, Haggman H (2004) Does extraction of DNA and RNA by magnetic fishing work for diverse plant species? Mol Biotechnol 27:209–215CrossRefGoogle Scholar
  54. Wahlberg J, Holmberg A, Bergh S, Hultman T, Uhlen M (1992) Automated magnetic preparation of DNA templates for solid phase sequencing. Electrophoresis 13:547–551CrossRefGoogle Scholar
  55. Watson RJ, Blackwell B (2000) Purification and characterization of a common soil component which inhibits the polymerase chain reaction. Can J Microbiol 46:633–642CrossRefGoogle Scholar
  56. Wilcockson J (1973) The use of sodium perchlorate in deproteinization during the preparation fsof nucleic acids. Biochem J 135:559–561Google Scholar
  57. Yoza B, Matsumoto M, Matsunaga T (2002) DNA extraction using modified bacterial magnetic particles in the presence of amino silane compound. J Biotechnol 94:217–224CrossRefGoogle Scholar
  58. Yoza B, Arakaki A, Matsunaga T (2003) DNA extraction using bacterial magnetic particles modified with hyperbranched polyamidoamine dendrimer. J Biotechnol 101:219-228CrossRefGoogle Scholar

Patent specifications and applications

  1. DE 100 13 995 A1 / EP 1 274 745 A1 Magnetische, silanisierte Trägermaterialien auf Basis von Polyvinylalkohol (2001) Parker J, Oster J, Brassard LÀ; chemagen Biopolymer-Technologie AG, GermanyGoogle Scholar
  2. DE 101 03 652 A1 Magnetische Polyvinylalkoholpartikel mit modifizierter Oberfläche zur Isolierung und Reinigung von Nukleinsäuren (2002) Brassard LÀ, Parker J, Smets H, Oster J; chemagen Biopolymer-Technologie AG, GermanyGoogle Scholar
  3. DE 41 43 639 C2 Verfahren zur Isolierung und Reinigung von Nukleinsäuren (1991) Colpan M; Qiagen GmbH, GermanyGoogle Scholar
  4. DE 41 27 657 A1 Perlförmige Polyvinylalkoholgele für die Aufreinigung und Auftrennung biologischer Flüssigkeiten, Verfahren zu ihrer Herstellung und Verwendung (1991) Müller-Schulte D; GermanyGoogle Scholar
  5. EP 0 003 905 B2 Process for preparing an aqueous emulsion or dispersion of a partly water-soluble material (1979) Ugelstad J; SINTEF, NorwayGoogle Scholar
  6. EP 0 728 198 B1 Isolation of nucleic acid (1994) Breivik J, Gaudernack G, Spurkland A; Qiagen GmbH, GermanyGoogle Scholar
  7. EP 0 796 327 B1 Isolation of nucleic acid (1995) Deggerdal AH, Larsen F; Dynal A/SGoogle Scholar
  8. EP 1 468 430 A1 Siliziumhaltige Magnetpartikel: Verfahren zu deren Herstellung und Verwendung der Partikel (2003) Hennig G, Hildenbrand K; Bayer AG, GermanyGoogle Scholar
  9. US 4,336,173 Process for preparing an aqueous emulsion or dispersion of a partly water-soluble material, and optionally further conversion of the prepared dispersion or emulsion to a polymer dispersion when the partly water-soluble material is a polymerizable monomer (1980) Ugelstad J; SINTEF, NorwayGoogle Scholar
  10. US 4,530,956 Process for the preparation of aqueous dispersions of organic materials and possible further conversion to a polymer dispersion when the organic material is a polymerizable monomer (1985) Ugelstad J, Berge A; SINTEF NorwayGoogle Scholar
  11. US 4,459,378 Monodisperse polymer particles and dispersions thereof (1982) Ugelstad J; SINTEF, NorwayGoogle Scholar
  12. US 5,075,430 Process for purification of DNA on diatomaceous earth (1990) Little MC.; Bio-Rad LaboratoriesGoogle Scholar
  13. US 5,512,439 Oligonucleotide-linked magnetic particles and uses thereof (1994) Hornes E, Korsnes L; Dynal AS, Oslo, NorwayGoogle Scholar
  14. US 5,523,231 Method to isolate macromolecules using magnetically attractable beads which do not specifically bind the macromolecules (1996) Reeve MA; Amersham International PLC, GBGoogle Scholar
  15. US 5,234,809 and EP 0 389 063 B1 Process for isolating nucleic acid (1991) Boom WR, Adriaanse HMA, Kievits T, Lens PF; Akzo N.V., the NetherlandsGoogle Scholar
  16. US 6,204,033 Preparation of polyvinyl alcohol-based magnetic particles for binding biomolecules (1998) Müller-Schulte D; GermanyGoogle Scholar
  17. US 6,514,688 Separating, detecting or quantifying biological materials using magnetic cross-linked polyvinyl alcohol particles (2000) Müller-Schultem D; chemagen Biopolymer-Technologie AG, GermanyGoogle Scholar
  18. WO/1991/012079 Method to isolate macromolecules using magnetically attractable beads which do not specifically bind the macromolecules (1991) Reeve MA; Amersham International PLCGoogle Scholar
  19. WO/1996/041811 Magnetic pigment (1996) Kleiber J, Walter T, Harttig H, Lesniak C, Mennig M, Riedling M, Schmidt H; Boehringer Mannheim GmbH, GermanyGoogle Scholar
  20. WO/1998/012717 Spherical magnetic particles (1996) Anselmann R, Pellatt MG; Merck Patent GmbH, GermanyGoogle Scholar
  21. WO/2002/009125 Spherical, magnetic SiO2 particles with an adjustable particle and pore size and an adjustable magnetic content. Method for producing them and use of SiO2 particles of this type (2001) Müller-Schulte D, Fischer R; Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. GermanyGoogle Scholar
  22. WO/2005/50581 A3 Sphärische, magnetische Silicagel-Träger mit vergrößerter Oberfläche für die Aufreinigung von Nukleinsäuren (2003) Müller-Schulte D; MagnaMedics GmbH, GermanyGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Forschungszentrum KarlsruheInstitute for Technical Chemistry, Water Technology and Geotechnology DivisionEggenstein-LeopoldshafenGermany

Personalised recommendations