Improved RNA extraction method using the BioMasher and BioMasher power-plus.
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.
Nippi Research Institute of Biomatrix, 520-11 Kuwabara, Toride, Ibaraki 302-0017, Japan. t-yamamoto@nippi-inc.co.jp
Takuji
T
Yamamoto
Kentaro
K
Nakashima
Yukio
Y
Maruta
Tomomi
T
Kiriyama
Michi
M
Sasaki
Shunpei
S
Sugiyama
Kana
K
Suzuki
Hitomi
H
Fujisaki
Jun
J
Sasaki
Yuko
Y
Kaku-Ushiki
Masatoshi
M
Tanida
Shinkichi
S
Irie
Shunji
S
Hattori
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D059018:DNA Contamination; D004587:Electrophoresis, Agar Gel; D005821:Genetic Techniques; D018551:Solanum lycopersicum; D051379:Mice; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D015203:Reproducibility of Results
D012313:RNA