Total RNA in the slices was isolated using ISOGEN® reagent (Nippon Gene, Tokyo, Japan). Gastric slices were homogenized using a bead homogenizer (Taitec, Saitama, Japan) with ISOGEN® reagent. The RNA was reverse-transcribed using ReverTra Ace® qPCR RT master mix with gDNA remover (Toyobo, Osaka, Japan). qPCR analysis was performed in duplicate using a CFX Connect™ real-time system (Bio-Rad Laboratories, Hercules, CA, USA) along with SYBR® premix Ex Taq™ II (Tli RNaseH Plus) (Takara Bio, Kusatsu, Japan). All gene-specific mRNA expression values were normalized to the expression value of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA. The primer sequences for each gene are listed in Supplementary Table S1 .
Bead homogenizer
Manufactured by TAITEC
Sourced in Japan
The Bead Homogenizer is a laboratory equipment used for the mechanical disruption and homogenization of biological samples, such as tissues, cells, and microorganisms. It utilizes rapid agitation of the sample in the presence of small beads to break down the sample into a homogeneous mixture.
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Lab products found in correlation
2 protocols using bead homogenizer
1
Comprehensive RNA Extraction and qPCR Analysis
2
Gastrocnemius Amino Acid Quantification
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The gastrocnemius muscle was homogenized in five volumes of ice-cold 5% 5-sulfosalicylic acid using a bead homogenizer (TAITEC) and centrifuged at 10,000×g for 10 min at 4 °C53 (link). Measurement of amino acid concentration in the supernatant was outsourced to Organization for Research Initiative and Promotion at Tottori University (Tottori, Japan) and measured using an automatic amino acid analyzer (JLC-500/V2, JEOL, Ltd., Tokyo, Japan).
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