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一种用于手动或自动纯化荧光标记核酸的快速方法,用于测序、基因分型和微阵列分析。

A rapid method for manual or automated purification of fluorescently labeled nucleic acids for sequencing, genotyping, and microarrays.

作者信息

Springer Amy L, Booth Lisa R, Braid Michael D, Houde Christiane M, Hughes Karin A, Kaiser Robert J, Pedrak Casandra, Spicer Douglas A, Stolyar Sergey

机构信息

Prolinx. Inc., Bothell, WA, USA.

出版信息

J Biomol Tech. 2003 Mar;14(1):17-32.

Abstract

Fluorescent dyes provide specific, sensitive, and multiplexed detection of nucleic acids. To maximize sensitivity, fluorescently labeled reaction products (e.g., cycle sequencing or primer extension products) must be purified away from residual dye-labeled precursors. Successful high-throughput analyses require that this purification be reliable, rapid, and amenable to automation. Common methods for purifying reaction products involve several steps and require processes that are not easily automated. Prolinx, Inc. has devel oped RapXtract superparamagnetic separation technology affording rapid and easy-to-perform methods that yield high-quality product and are easily automated. The technology uses superparamagnetic particles that specifically remove unincorporated dye-labeled precursors. These particles are efficiently pelleted in the presence of a magnetic field, making them ideal for purification because of the rapid separations that they allow. RapXtract-purified sequencing reactions yield data with good signal and high Phred quality scores, and they work with various sequencing dye chemistries, including BigDye and near-infrared fluorescence IRDyes. RapXtract technology can also be used to purify dye primer sequencing reactions, primer extension reactions for genotyping analysis, and nucleic acid labeling reactions for microarray hybridization. The ease of use and versatility of RapXtract technology makes it a good choice for manual or automated purification of fluorescently labeled nucleic acids.

摘要

荧光染料可对核酸进行特异性、灵敏且多重的检测。为了将灵敏度最大化,必须将荧光标记的反应产物(例如循环测序或引物延伸产物)与残留的染料标记前体分离并纯化。成功的高通量分析要求这种纯化过程可靠、快速且易于自动化。常见的反应产物纯化方法涉及多个步骤,且所需过程不易自动化。Prolinx公司开发了RapXtract超顺磁分离技术,该技术提供了快速且易于操作的方法,能够产生高质量的产物并且易于自动化。该技术使用超顺磁颗粒特异性去除未掺入的染料标记前体。这些颗粒在磁场存在下能有效沉淀,因其能实现快速分离,所以非常适合用于纯化。经RapXtract纯化的测序反应能产生具有良好信号和高Phred质量分数的数据,并且适用于各种测序染料化学体系,包括BigDye和近红外荧光IRDyes。RapXtract技术还可用于纯化染料引物测序反应、用于基因分型分析的引物延伸反应以及用于微阵列杂交的核酸标记反应。RapXtract技术的易用性和多功能性使其成为手动或自动纯化荧光标记核酸的理想选择。

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