African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Review

Whole genome amplification: Use of advanced isothermal method

  Sima Moghaddaszadeh Ahrabi1, Safar Farajnia2,3*, Ghodratollah Rahimi-Mianji4, Soheila Montazer Saheb3, Ardeshir Nejati-Javaremi5  
  1Ph.D. Student, Department of Animal Science, Science and Research Branch, Islamic Azad University(IAU), Tehran, Iran. 2Biotechnology Researches Center, Tabriz University of Medical Sciences, Tabriz, Iran. 3Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 4Department of Animal Improvement and Genetics, Sari Agricultural Sciences and Natural Resources University, Iran. 5Department of Animal Science, College of Agriculture & Natural Resources, University of Tehran, Karaj Campus, Iran.  
Email: [email protected]

  •  Accepted: 23 December 2010
  •  Published: 29 December 2010

Abstract

 

Laboratory method for amplifying genomic deoxyribonucleic acid (DNA) samples, aiming to generate more amounts and sufficient quantity DNA for further subsequent specific analysis is named, whole genome amplification (WGA). This method is only a way to increase input material from few cells and limited DNA contents. While polymerase chain reaction (PCR)-based WGA methods have been under continuous development for over a decade, shortcomings of these methods enforced many researchers to become switch to the use of non-PCR-based linear amplification techniques. Moreover, application of one high fidelity and high possessive DNA polymerase enabled an isothermal WGA technique named, multiple displacement amplification (MDA). MDA is not based on PCR and does not require thermal cycling. It should be noted that, while MDA-based techniques proposed aiming to overcome the drawbacks of PCR-based methods however, MDA is still facing some challenges. It seems that PCR-based WGA methods also have some merits. One of the problems which encountered both MDA and PCR-based method is in the amplification degraded DNA templates. WGA methods such as T7-based linear amplification of DNA (TLAD), balanced-PCR amplification, and restriction and circulation-aided rolling circle amplification (RCA-RCA) have been suggested to aim at amplification of such DNA templates.

 

Key words: Whole genome amplification, multiple displacement amplification (MDA), non PCR-based methods.