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首页> 外文期刊>BMC Genomics >Primer fabrication using polymerase mediated oligonucleotide synthesis
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Primer fabrication using polymerase mediated oligonucleotide synthesis

机译：使用聚合酶介导的寡核苷酸合成进行引物制备

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Background Custom solid phase oligonucleotide synthesis is an important foundation supporting nearly every aspect of current genomics. In spite of the demand for oligonucleotide primers, their synthesis remains relatively expensive, time consuming and in many circumstances a wasteful process. In this methodology, described as polymerase mediated oligonucleotide synthesis (PMOS), a DNA polymerase is used to increase the hybridization affinity of one oligonucleotide by using another as a template for DNA synthesis. This self-assembly process provides an opportunity to instantly generate a very large number of useful gene-specific primers from a small library of simple precursors. PMOS can be used to generate primers directly in the end-users laboratory within the context of any DNA polymerase chemistry such as in PCR or sequencing reactions Results To demonstrate the utility of PMOS, a universal 768-member oligonucleotide library (UniSeq) was designed, fabricated and its performance optimized and evaluated in a range of PCR and DNA sequencing reactions. This methodology used to derive specific 11-mers, performed well in each of these activities and produced the desired amplification or sequencing analysis with results comparable to primers made by time consuming and expensive custom synthesis. Conclusion On the basis of these experiments, we believe this novel system would be broadly applicable and could in many circumstances replace the need for conventional oligonucleotide synthesis.
机译：背景技术定制固相寡核苷酸合成是支持当前基因组学几乎每个方面的重要基础。尽管需要寡核苷酸引物，但是它们的合成仍然相对昂贵，费时并且在许多情况下是浪费的过程。在这种称为聚合酶介导的寡核苷酸合成（PMOS）的方法中，DNA聚合酶用于通过将另一种寡核苷酸用作DNA合成模板来增加一种寡核苷酸的杂交亲和力。这种自组装过程提供了一个机会，可以从一个小的简单前体文库中立即生成大量有用的基因特异性引物。在任何DNA聚合酶化学方法（例如PCR或测序反应）的背景下，PMOS均可用于在最终用户实验室中直接生成引物。结果为了证明PMOS的实用性，设计了通用768元寡核苷酸文库（UniSeq），可以在一系列PCR和DNA测序反应中进行优化和评估。该方法用于衍生特定的11-聚体，在这些活性的每一种中均表现良好，并产生了所需的扩增或测序分析，其结果可与耗时且昂贵的定制合成制备的引物相媲美。结论基于这些实验，我们认为该新型系统将广泛适用，并在许多情况下可以取代常规寡核苷酸合成的需求。

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