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首页> 外文期刊>The Journal of molecular diagnostics: JMD >Immiscible phase nucleic acid purification eliminates PCR inhibitors with a single pass of paramagnetic particles through a hydrophobic liquid.
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Immiscible phase nucleic acid purification eliminates PCR inhibitors with a single pass of paramagnetic particles through a hydrophobic liquid.

机译：互溶相核酸纯化使顺磁性颗粒单次通过疏水性液体即可消除PCR抑制剂。

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摘要

Extraction and purification of nucleic acids from complex biological samples for PCR are critical steps because inhibitors must be removed that can affect reaction efficiency and the accuracy of results. This preanalytical processing generally involves capturing nucleic acids on microparticles that are then washed with a series of buffers to desorb and dilute out interfering substances. We have developed a novel purification method that replaces multiple wash steps with a single pass of paramagnetic particles (PMPs) though an immiscible hydrophobic liquid. Only two aqueous solutions are required: a lysis buffer, in which nucleic acids are captured on PMPs, and an elution buffer, in which they are released for amplification. The PMPs containing the nucleic acids are magnetically transported through a channel containing liquid wax that connects the lysis chamber to the elution chamber in a specially designed cartridge. Transporting PMPs through the immiscible phase yielded DNA and RNA as pure as that obtained after extensive wash steps required by comparable purification methods. Our immiscible-phase process has been applied to targets in whole blood, plasma, and urine and will enable the development of faster and simpler purification systems.
机译：从复杂的生物样品中提取和纯化核酸用于PCR是关键步骤，因为必须去除会影响反应效率和结果准确性的抑制剂。这种分析前的处理通常涉及在微粒上捕获核酸，然后用一系列缓冲液洗涤核酸以解吸和稀释干扰物质。我们已经开发出一种新颖的纯化方法，该方法通过不混溶的疏水性液体通过单次通过的顺磁性颗粒（PMP）代替了多个洗涤步骤。仅需要两种水溶液：一种裂解缓冲液（其中核酸被捕获在PMP上）和一种洗脱缓冲液（其中它们被释放以进行扩增）。包含核酸的PMP通过一个包含液态蜡的通道进行磁性传输，该通道将裂解室连接到专门设计的盒中的洗脱室。通过不溶混相运输PMP产生的DNA和RNA纯度与可比的纯化方法所需的大量洗涤步骤后的纯度相同。我们的不相溶过程已应用于全血，血浆和尿液中的目标，将能够开发更快，更简单的纯化系统。

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