Whole genome amplification: Use of advanced isothermal method

Abstract

Laboratory method for amplifying genomic deoxyribonucleic acid (DNA) samples aiming to generate more amounts and sufficient quantity DNA for subsequent specific analysis is named whole genome amplification (WGA). This method is only way to increase input material from few cells and limited DNA contents. While PCR-based WGA methods have been under continuous development for over a decade, shortcomings of these methods enforced many researchers to switch to the use of non-PCR-based linear amplification techniques. Moreover, application of high fidelity and high possessive DNA polymerases enabled development of an isothermal WGA technique named multiple displacement amplification (MDA). MDA is not based on PCR and doses 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 methods is in the amplification of degraded DNA templates. WGA methods such as T7-based linear amplification of DNA (TLAD), balanced-PCR amplification and restriction and circularization-aided rolling circle amplification (RCA-RCA) have been suggested to aim at amplification of such DNA templates.