Improved RNA Extraction Method Using the BioMasher and BioMasher Power-Plus

均质机 核糖核酸 RNA提取 均质化(气候) 萃取(化学) 色谱法 琼脂糖凝胶电泳 琼脂糖 生物 分子生物学 化学 生物化学 DNA 生态学 基因 生物多样性
作者
Takuji Yamamoto,Kentaro Nakashima,Yukio Maruta,Tomomi Kiriyama,Michi Sasaki,Shunpei Sugiyama,Kana Suzuki,Hitomi Fujisaki,Jun Sasaki,Yuko Kaku-Ushiki,Masatoshi Tanida,Shinkichi Irie,Shunji Hattori
出处
期刊:Journal of Veterinary Medical Science [Japanese Society of Veterinary Science]
卷期号:74 (12): 1561-1567 被引量:6
标识
DOI:10.1292/jvms.12-0213
摘要

The BioMasher is a disposable homogenizer that was developed to homogenize bovine brain tissue for bovine spongiform encephalopathy diagnosis. Capable of preventing the biohazard risk from infectious samples, it also prevents cross-contamination among samples. The BioMasher is thus widely used in biochemical research, especially for RNA extraction. Here, we tested a novel BioMasher application for RNA extraction from animal and plant tissues. We also developed a grinding machine specific for the BioMasher, named the BioMasher Power-Plus. We developed RNA extraction protocols using the BioMasher combined with the BioMasher Power-Plus. We compared RNA extraction efficiency of the BioMasher with that of the FastPrep and the glass homogenizer. Though the RNA extraction efficiency by the BioMasher was nearly equivalent to that of the FastPrep and the glass homogenizer, sample preparation time was shorter for the BioMasher. The utility of RNA extraction by the BioMasher was examined in mouse, rat, and tomato tissue samples. In the rodent tissues, the highest extraction efficiency of total RNA was from liver, with lowest efficiency from fibrous tissues such as muscle. The quality of extracted total RNA was confirmed by agarose gel electrophoresis which produced highly visible clear bands of 18S and 28S rRNAs. Reproducibility among different operators in RNA extraction from tomato roots was improved by using the BioMasher Power-Plus. The BioMasher and BioMasher Power-Plus provide an effective and easy homogenization method for total RNA extraction from some rodent and plant tissues.

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