One gram of fecal sample was weighed and homogenized in 40 mL of PBS with a benchtop homogenizer (MP Biomedical). The sample was then centrifuged at 17,000x g for 5 min and the supernatant was filtered sequentially through a 2 μm and 0.45 μm filter. Phages were then concentrated using a polyethylene glycol (PEG) method(Luong et al., 2020 (link)). Briefly, one molar solid NaCl and 10% (v/v) PEG 8000 (Sigma) were dissolved in PBS and incubated overnight at 4°C as recommended for a constant and stable precipitation. The phages mixed in the solution were pelleted by centrifugation at 5,250x g for 1 h at 4°C and re-suspended in 500 μl of SM buffer (NaCl 100 mM, MgSO4.7H2O 8 mM, Tris-Cl 50 mM).
Benchtop homogenizer
Manufactured by MP Biomedicals
Sourced in United States
The Benchtop Homogenizer is a versatile laboratory equipment designed for efficient sample preparation. It is used to disrupt and homogenize a wide range of materials, including tissues, cells, and other solid or semisolid samples. The device's compact design makes it suitable for use on a laboratory benchtop.
Automatically generated - may contain errors
2 protocols using benchtop homogenizer
1
Phage Extraction from Fecal Samples
2
RNA Extraction from Cell Pellets
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Cultures from 3 ml samples incubated under the condition above were harvested by centrifugation. Cell pellets (~ 1 × 109 cells) were suspended in 1 ml Trizol Reagent and disrupted with benchtop homogenizer (MP Biomedicals, California, USA). Then total RNA was extracted and the quantity of RNA was determined by a NanoDrop spectrophotometer (ThermoFisher Scientific, Waltham, MA). The quality of RNA was assayed using an Agilent 2100 bioanalyzer (Agilent Technologies, Waldbronn, Germany).
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