Murine fecal samples were mixed with 95% ethanol (Nacalai Tesque) to a concentration of 100 mg/ml and homogenized by using two 15-s pulses at 6500 rpm from a tissue homogenizer (Precellys 24, Bertin Instruments, Montigny-le-Bretonneux, France) with zirconia beads (Tomy Digital Biology Co., Ltd, Tokyo, Japan). The homogenate was centrifuged at 1600 × g for 10 min at 4 °C and the supernatant collected. The fecal supernatant or bacterial cultured medium were labeled using an FA Labeling kit (YMC Co., Ltd, Kyoto, Japan) in accordance with the manufacturer’s instructions. The labeled samples were analyzed on an HPLC system (Ultimate 3000, Thermo Fisher Scientific, Waltham, Massachusetts, USA) with a 6.0 × 250 mm YMC-Pack FA column (YMC), and the UV spectrum at 400 nm was measured.
Zirconia beads
Manufactured by TOMY
Sourced in France, Japan
Zirconia beads are a type of lab equipment used for grinding and milling applications. They are made of zirconium oxide, a hard and durable ceramic material. Zirconia beads are commonly used in the milling and homogenization of samples in various industries, including pharmaceutical, chemical, and material science research.
Automatically generated - may contain errors
Lab products found in correlation
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Pack fa column, by YMC (1 mentions)
Precellys 24, by Bertin Technologies (1 mentions)
Micro smash, by TOMY (1 mentions)
Bead ruptor elite, by Omni International (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
3 protocols using zirconia beads
1
Fecal Fatty Acid Profiling
2
Isolation of Matsutake Mycorrhizal Fungi
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Matsutake ectomycorrhizal root tips were picked from shiro soil and from the seedlings of P. densiflora under a stereomicroscope. The root tips, 3-5 mm length, were rinsed five times with sterile water followed by stirring in a beaker first in 0.05% Tween 20 for 2 min and then in 0.001% Tween 20 for 30 min. Twenty mycorrhizal root tips per shiro sample were collected into a 2 ml tube with Zirconia beads, 2.0 mm (TOMY, Japan). Warm HNC-medium (200 μl) containing yeast extract (60 g), CaCl 2 (0.5 g), SDS (5 g) in 1 L water (Nonomura and Hayakawa 1988) was added. The root tissues were disrupted using a Micro Smash ™ (TOMY, MS-100, Japan) with 20 s pulses at 2000 rpm, repeated three times, then incubated at 42 °C for 30 min. The tubes were centrifuged at 8000 rpm for 2 min. The supernatant was looped by a glass loop onto ISP2 agar medium (Shirling and Gottlieb 1966) (link) surface, following a distinctive zig-zag pattern to ensure obtaining a single colony. The cultures were incubated at 28 °C in the dark. Later, a single colony was transferred onto a fresh ISP2 medium in the same conditions.
3
RNA Isolation from Ketamine-Xylazine Anesthetized Mice
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Ketamine xylazine–anesthetized mice (150 mg/kg of ketamine, 10 mg/kg of xylazine) were intracardially perfused with PBS and organs were collected in tubes containing zirconia beads (TOMY Digital Biology) and 1 ml TRIzol (Invitrogen) and stored at −80°C. To isolate RNA, tissues were thawed on ice and homogenized using the Bead Ruptor Elite (OMNI International) at a speed of 6 m/s (3 cycles of lysis × 30 s). Tissue lysates were then transferred to 1.5-ml Eppendorf tubes and mixed with 230 μl of chloroform and incubated at RT for 2 min. Samples were centrifuged at 12,000g for 10 min at 4°C and recovered supernatants were transferred into clean Eppendorf Tubes and mixed with 70% ethanol (1:1 volume). RNA was purified using the RNeasy Mini Kit (QIAGEN) according to the manufacturer’s protocol. Viral RNA copies were quantified by qRT-PCR as described above.
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