Multidisplacement amplification

The multiple-displacement amplification (MDA), also beach-displacement amplification (SDA) or isothermal multiple-displacement amplification (IMDA) is a method for the reproduction of small amounts of DNA as a starting material. It is a variant of isothermal DNA amplification .

properties

Compared to the polymerase chain reaction , this method is particularly suitable for the replication of long DNA sequences (more than 2,000 to 100,000 base pairs). Another advantage is that with mixtures of DNA there is no bias due to this type of amplification . This means that in the case of mixtures, individual DNA sequences are not preferred or others are disadvantaged.

Due to these properties, this method is particularly suitable for the propagation of z. B. highly complex mixtures of genomic DNA. A few nanograms are sufficient as the starting material (e.g. the DNA of a few hundred cells), which in MDA is multiplied about 1,000 to 10,000 times with the help of DNA polymerase .

principle

In comparison to the polymerase chain reaction, the MDA is carried out at a constant temperature. First, the double-stranded template DNA is separated into single strands by means of denaturation. Then the Φ29 DNA polymerase and a mixture of deoxynucleotides are added. The Φ29 DNA polymerase attaches itself to the single strands of the DNA and then produces the second complementary strand. The complementary strand is lengthened about 25 to 50 bases per second with an error rate of 1 error to 3 × 10 ⁶ bases. In comparison: with a conventional PCR with Taq polymerase one calculates with a speed of 1000 to 2000 bases per minute and an error rate of 1 error per 2 × 10 ³ bases. Alternative isothermal methods for generating new DNA sequences are e.g. B. isothermal assembly and circular polymerase extension cloning (CPEC).

Alternative methods of amplifying DNA are e.g. B. the polymerase chain reaction , the nicking enzyme amplification reaction , the isothermal assembly , the loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification ( NASBA ), the helicase-dependent amplification (HDA), the recombinase polymerase amplification and the rolling circle replication (RCA). Further detection methods are e.g. B. Nicking endonuclease signal amplification ( NESA ) and nicking endonuclease assisted nanoparticle activation ( NENNA ), exonuclease-aided target recycling , junction or Y-probes , split DNAZyme and deoxyribozyme amplification , non-covalent DNA catalysis and the hybridization chain reaction ( HCR).

literature

Rajyalakshmi Luthra, L. Jeffrey Medeiros: Isothermal Multiple Displacement Amplification - A Highly Reliable Approach for Generating Unlimited High Molecular Weight Genomic DNA from Clinical Specimens. In: J. Mol. Diagn. Vol. 6, No. 3, 2004, pp. 236-242, PMID 15269301 .
Frank B. Dean et al. : Comprehensive human genome amplification using multiple displacement amplification. In: Proc. Natl. Acad. Sci. USA Vol. 99, No. 8, 2002, pp. 5261-5266, PMID 11959976 .
Pooria Gill, Amir Ghaemi: Nucleic acid isothermal amplification technologies: a review. In: Nucleosides Nucleotides Nucleic Acids Vol. 27, No. 3, 2008, pp. 224-43, PMID 18260008 .

Web links

Gerald Schock, Nina Gildehaus, Helge Lubenow, Dirk Löffert and Christian Korfhage: Unlimited DNA analyzes through multiple displacement amplification ; biospektrum, 6/2005

Individual evidence

↑ L. Westin, X. Xu, C. Miller, L. Wang, CF Edman, M. Nerenberg: Anchored multiplex amplification on a microelectronic chip array. In: Nature biotechnology . Volume 18, Number 2, February 2000, pp. 199-204, doi: 10.1038 / 72658 , PMID 10657128 .
↑ J. Quan, J. Tian: Circular polymerase extension cloning of complex gene libraries and pathways. In: PLOS ONE . Volume 4, number 7, 2009, p. E6441, doi: 10.1371 / journal.pone.0006441 , PMID 19649325 , PMC 2713398 (free full text).
↑ EC Oriero, J. Jacobs, JP Van Geertruyden, D. Nwakanma, U. D'Alessandro: Molecular-based isothermal tests for field diagnosis of malaria and their potential contribution to malaria elimination. In: Journal of Antimicrobial Chemotherapy . [Electronic publication before going to press] September 2014, doi: 10.1093 / jac / dku343 , PMID 25223973 .

↑ L. Yan, J. Zhou, Y. Zheng, AS Gamson, BT Roembke, S. Nakayama, HO Sintim: Isothermal amplified detection of DNA and RNA. In: Molecular bioSystems. Volume 10, Number 5, May 2014, ISSN 1742-2051 , pp. 970-1003, doi: 10.1039 / c3mb70304e , PMID 24643211 .

[1] L. Westin, X. Xu, C. Miller, L. Wang, CF Edman, M. Nerenberg: Anchored multiplex amplification on a microelectronic chip array. In: Nature biotechnology . Volume 18, Number 2, February 2000, pp. 199-204, doi: 10.1038 / 72658 , PMID 10657128 .

[2] J. Quan, J. Tian: Circular polymerase extension cloning of complex gene libraries and pathways. In: PLOS ONE . Volume 4, number 7, 2009, p. E6441, doi: 10.1371 / journal.pone.0006441 , PMID 19649325 , PMC 2713398 (free full text).

[3] EC Oriero, J. Jacobs, JP Van Geertruyden, D. Nwakanma, U. D'Alessandro: Molecular-based isothermal tests for field diagnosis of malaria and their potential contribution to malaria elimination. In: Journal of Antimicrobial Chemotherapy . [Electronic publication before going to press] September 2014, doi: 10.1093 / jac / dku343 , PMID 25223973 .

[4] L. Yan, J. Zhou, Y. Zheng, AS Gamson, BT Roembke, S. Nakayama, HO Sintim: Isothermal amplified detection of DNA and RNA. In: Molecular bioSystems. Volume 10, Number 5, May 2014, ISSN 1742-2051 , pp. 970-1003, doi: 10.1039 / c3mb70304e , PMID 24643211 .