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首页> 外文期刊>Acta Biochimica Polonica >Principles and applications of Ligation Mediated PCR methods for DNA-based typing of microbial organisms
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Principles and applications of Ligation Mediated PCR methods for DNA-based typing of microbial organisms

机译：连接介导PCR方法在基于DNA的微生物分类中的原理和应用

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摘要

A significant number of DNA-based techniques has been introduced into the field of microorganisms' characterization and taxonomy. These genomic fingerprinting methods were developed to detect DNA sequence polymorphisms by using general principles, such as restriction endonuclease analysis, molecular hybridization, and PCR amplification. In recent years, some alternative techniques based on ligation of oligonucleotide adapters before DNA amplification by PCR, known as Ligation-Mediated PCR methods ( LM PCR), have been successfully applied for the typing of microorganisms below the species level. These molecular methods include: Amplified Fragment Length Polymorphism ( AFLP), Amplification of DNA fragments Surrounding Rare Restriction Sites ( ADSRRS), PCR Melting Profiles ( PCR MP), Ligation Mediated PCR/Shifter ( LM PCR/Shifter), Infrequent-Restriction-Site Amplification ( IRS PCR), double digestion Ligation Mediated Suppression PCR ( ddLMS PCR). These techniques are now applied more and more often because they involve less time, are comparably inexpensive, and require only standard lab equipment. Here, we present a general review of this group of methods showing their possibilities and limitations. We also identify questions and propose solutions which may be helpful in choosing a particular LM PCR method for the achievement of the required goal.
机译：大量基于DNA的技术已被引入微生物的表征和分类学领域。这些基因组指纹图谱方法是通过使用一般原理（例如限制性核酸内切酶分析，分子杂交和PCR扩增）来检测DNA序列多态性的。近年来，一些基于寡核苷酸衔接子在PCR扩增之前进行连接的替代技术，被称为连接介导PCR方法（LM PCR），已成功应用于物种水平以下的微生物分型。这些分子方法包括：扩增片段长度多态性（AFLP），稀有限制位点周围DNA片段的扩增（ADSRRS），PCR熔解谱（PCR MP），连接介导的PCR /移位器（LM PCR / Shifter），不频繁限制位点扩增（IRS PCR），双酶切连接介导的抑制PCR（ddLMS PCR）。这些技术现在越来越多地被应用，因为它们占用的时间更少，相对便宜且仅需要标准实验室设备。在这里，我们对这组方法进行了概述，显示了它们的可能性和局限性。我们还将找出问题并提出解决方案，这可能有助于选择特定的LM PCR方法来实现所需的目标。

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