Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification
Electroporation and lysis of marine microalga Karenia brevis for RNA extraction and amplification
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DOI：
10.1098/rsif.2010.0445
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发表时间：
2011-04-01
影响因子：
3.9
通讯作者：
Morgan, H.
中科院分区：
文献类型：
--
作者：
Bahi, M. M.;Tsaloglou, M. -N.;Morgan, H.
我们在这里描述了一个简单的装置介电电泳浓缩海洋microarenia brevis nonmotile细胞，然后通过电场介导的裂解RNA提取。使用磁珠纯化裂解物并提取纯RNA。通过基于核酸序列的扩增在芯片外评估RNA质量，并确定裂解的最佳条件。该程序将形成集成微流体系统的一部分，该系统正在开发用于进行细胞浓缩和裂解、RNA提取/纯化和实时定量RNA检测的子系统。该集成系统及其组件可用于广泛的应用，包括原位有害藻华检测、转录组学和即时诊断。
We describe here a simple device for dielectrophoretic concentration of marine microalga Karenia brevis nonmotile cells, followed by electric field-mediated lysis for RNA extraction. The lysate was purified using magnetic beads and pure RNA extracted. RNA quality was assessed off-chip by nucleic acid sequence-based amplification and the optimum conditions for lysis were determined. This procedure will form part of an integrated microfluidic system that is being developed with sub-systems for performing cell concentration and lysis, RNA extraction/purification and real-time quantitative RNA detection. The integrated system and its components could be used for a large range of applications including in situ harmful algal bloom detection, transcriptomics and point-of-care diagnostics.