RNA chip: quality assessment of RNA by microchannel linear gel electrophoresis in injection-molded plastic chips.
RNA chip: quality assessment of RNA by microchannel linear gel electrophoresis in injection-molded plastic chips.
复制标题
RNA 芯片：通过注塑塑料芯片中的微通道线性凝胶电泳对 RNA 进行质量评估。
作者：
M. Ogura;Yoichi Agata;Watanabe Kenji;R. McCormick;Y. Hamaguchi;Yoshimasa Aso;M. Mitsuhashi;M. Mitsuhashi
rRNA的两种主要成分（18 S和28 S rRNA）通过电泳在注塑成型的丙烯酸芯片中分离，该芯片具有宽100 μ m、深40 μ m的微通道和1 cm的分离距离。微通道填充有4g/L羟丙基甲基纤维素作为筛分聚合物和5 mg/L溴化乙锭用于RNA染色。通过配备有光度计和590 nm发射滤光片的荧光显微镜检测荧光信号。该测定快速（<3分钟），可重复，不含RNase，仅需1-2微升样品。检测限约为10 mg/L（10 ng/μ L），比常规琼脂糖凝胶电泳低100倍。因为仅使用0.1 nL的上样样品进行电泳，所以分离中rRNA的可检测峰来源于比单个细胞中更少的RNA。由于RNA的质量对于RNA相关的诊断测试至关重要，因此一次性塑料芯片将用于RNA的质量评估。
Two major components of rRNA (18S and 28S rRNA) were separated by electrophoresis in injection-molded acrylic chips with a microchannel 100 microm in width, 40 microm in depth, and with 1 cm of separation distance. Microchannels were filled with 4 g/L hydroxypropylmethylcellulose as sieving polymer and 5 mg/L ethidium bromide for RNA staining. The fluorescent signals were detected by a fluorescent microscope equipped with a photometer and 590 nm emission filter. The assay is rapid (<3 min), reproducible, RNase-free, and requires only 1-2 microL of sample. The detection limit was approximately 10 mg/L (10 ng/microL), 100-fold lower than that for conventional agarose gel electrophoresis. Because only 0.1 nL of the loaded sample was used for electrophoresis, the detectable peaks of rRNA in the separation were derived from less RNA than in a single cell. Because the quality of RNA is critical for RNA-related diagnostic tests, disposable plastic chips will be useful for quality assessment of RNA.