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在基于探针的定量聚合酶链反应中使用合成寡核苷酸作为标准品。

Using synthetic oligonucleotides as standards in probe-based qPCR.

作者信息

Conte Jillian, Potoczniak Margret J, Tobe Shanan S

机构信息

Keystone College, Department of Biological & Physical Sciences, One College Green, La Plume, PA 18440, USA.

University of the Sciences, Department of Biological Sciences, Philadelphia, PA, USA.

出版信息

Biotechniques. 2018 Apr;64(4):177-179. doi: 10.2144/btn-2018-2000.

Abstract

Real-time PCR (qPCR) is widely used in the life sciences. For quantifying DNA, a standard curve is required. Common methods for standard development are time consuming, costly, necessitate a specific skill set, and pose a contamination risk. Using a targeted synthetic oligonucleotide, such as a gBlocks Gene Fragment, overcomes these drawbacks and provides researchers an accurate and quick solution to standard development. Here, we demonstrate that using a gBlocks fragment as a standard provides comparable sensitivity, reliability, and assay performance to a purified amplicon standard.

摘要

实时定量聚合酶链反应(qPCR)在生命科学领域被广泛应用。为了对DNA进行定量,需要构建标准曲线。传统的标准曲线构建方法耗时、成本高、需要特定的技能,并且存在污染风险。使用靶向合成寡核苷酸,如gBlocks基因片段,可以克服这些缺点,并为研究人员提供一种准确、快速的标准曲线构建解决方案。在此,我们证明使用gBlocks片段作为标准物,其灵敏度、可靠性和检测性能与纯化的扩增子标准物相当。

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