Department of Nano Engineering, Center for Nano Manufacturing, Inje University, 607 Obang-dong, Gimhae, GyongNam, 621-749, Republic of Korea.
Biomed Microdevices. 2013 Feb;15(1):9-15. doi: 10.1007/s10544-012-9682-3.
This report introduces an on-chip integrated reverse transcription (RT)-microchip, which includes two genetic functionalities of RNA extraction and cDNA synthesis. In the RNA extraction compartment, RNA is extracted from peripheral blood lysate within 1 min, by lateral magnetophoresis using magnetic oligo-dT beads. The extracted RNA is then collected and used directly to produce cDNA in the cDNA synthesis microchamber, which is monolithically integrated with the RNA extraction compartment. To verify the superiority of the proposed RT-microchip, RT-PCR amplification was performed using cDNA harvested from the RT-microchip, and the results were compared with those obtained using typical RNA extraction methods such as a silica matrix column and magnetic oligo-dT beads. The RT-PCR amplification results using 100 μl of blood showed that the intensity of the bands in gel electrophoresis of the RT-microchip was 2-fold stronger than that of the silica matrix column and 2.65-fold stronger than that of the magnetic oligo-dT beads. The results demonstrate that the RT-microchip technique is the most sensitive of the tested methods.
本报告介绍了一种片上集成的逆转录（RT）微芯片，它包括 RNA 提取和 cDNA 合成两个遗传功能。在 RNA 提取隔室中，通过使用磁性寡聚 dT 珠的横向磁泳，在 1 分钟内从外周血裂解液中提取 RNA。提取的 RNA 随后被收集并直接用于 cDNA 合成微室中的 cDNA 合成，该微室与 RNA 提取隔室整体集成。为了验证所提出的 RT 微芯片的优越性，使用从 RT 微芯片中收获的 cDNA 进行 RT-PCR 扩增，并将结果与使用典型的 RNA 提取方法（如硅基质柱和磁性寡聚 dT 珠）获得的结果进行比较。使用 100μl 血液进行的 RT-PCR 扩增结果表明，凝胶电泳中 RT 微芯片的条带强度比硅基质柱强 2 倍，比磁性寡聚 dT 珠强 2.65 倍。结果表明，RT 微芯片技术是测试方法中最敏感的。
