School of Nano Engineering, Inje University, 607 Obang-dong, Gimhae, GyongNam, 621-749, Republic of Korea.
Lab Chip. 2010 Oct 21;10(20):2764-70. doi: 10.1039/c005145d. Epub 2010 Sep 2.
This paper presents a high-speed RNA microextractor for the direct isolation of RNA from peripheral blood lysate using magnetic oligo-dT beads. The extraction is achieved through lateral magnetophoresis, generated by a ferromagnetic wire array inlaid on a glass substrate. This RNA microextractor separated more than 80% of magnetic beads with a flow rate up to 20 ml h(-1), and the overall extraction procedure was completed within 1 min. The absorbance ratio of RNA to protein (A(260)/A(280)) was >1.7, indicating that the extraction technology yielded nearly pure RNA. The feasibility of this technique was evaluated further for its applicability to reverse transcription polymerase chain reaction (RT-PCR) procedures by performing cDNA synthesis and PCR. The analysis verified that the RNA microextractor is a practical method for easy, rapid, and high-precision RT-PCR using minimal reagent volumes without requiring highly trained personnel. In addition, it can be readily incorporated into genetic analysis procedures for realizing automated on-chip genetic platforms in a micro format.
本文提出了一种从外周血裂解液中直接提取 RNA 的高速 RNA 微提取器，该提取器使用磁性寡聚 dT 珠。通过镶嵌在玻璃基板上的铁磁线阵列产生横向磁泳，实现了提取。这种 RNA 微提取器在流速高达 20ml/h 时可分离超过 80%的磁珠，整个提取过程在 1 分钟内完成。RNA 与蛋白质的吸光度比值（A260/A280）大于 1.7，表明提取技术得到了几乎纯净的 RNA。通过进行 cDNA 合成和 PCR，进一步评估了该技术用于逆转录聚合酶链反应（RT-PCR）程序的可行性。分析验证了 RNA 微提取器是一种实用的方法，可用于使用最小体积的试剂轻松、快速、高精度地进行 RT-PCR，而无需高度训练的人员。此外，它可以很容易地整合到遗传分析程序中，以实现微格式的自动化芯片遗传平台。
