Approximately 30 min, 6, 24 and 48 h after i.n. infection, mice were anaesthetised with 2.5 % (v/v) fluothane, and blood was collected by cardiac puncture. Mice were culled, and lungs were collected into 10 ml PBS and homogenised using an Ultra-Turrax T8 homogeniser (IKA-Werke, Germany). After i.v. infection, blood samples were collected from a tail vein at 24, 48 and 72 h. Viable counts from each homogenate and blood sample were determined by serial dilution using blood agar (Oxoid) containing 5 % (v/v) defibrinated horse blood and overnight incubation in an anaerobic jar at 37 °C. The colonies showed typical morphology for S. pneumoniae and α-hemolysis.
Ultra turrax t8 homogeniser
Manufactured by IKA Group
Sourced in Germany
The Ultra-Turrax T8 is a high-speed homogenizer designed for efficient mixing, dispersing, and emulsifying of various materials. It features a powerful motor and a range of dispersing elements to handle a variety of applications. The core function of the Ultra-Turrax T8 is to provide effective and consistent homogenization of samples.
Automatically generated - may contain errors
2 protocols using ultra turrax t8 homogeniser
1
Pneumococcal Infection Kinetics in Mice
2
RNA Extraction with TRIZOL Reagent
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RNA was extracted with TRIZOL (Invitrogen, UK) according to manufacturer's instruction. Briefly, 1 mL TRIZOL reagent per 50-100 mg of tissue sample was homogenised with Ultra-Turrax T8 homogeniser (IKA-Werke, Germany) in a type II flow hood (Gelaire, Flow Laboratory), and phase separated with 0.2 mL of chloroform per 1 mL of TRIZOL. RNA was precipitated with 0.5 mL of 100% isopropanol per 1 mL of TRIZOL reagent. RNA concentration was normalised to wet weight, and purity assessed by measuring UV 260/280 absorbance ratios (NanoDrop® ND-1000 UV-Vis Spectrophotometer). RNA quality was assessed by measuring RNA Integrity Number (RIN), rRNA ratio and microcapillary electrophoresis with Agilent Bioanalyzer 2100 with RNA 6000 Nano Chip (Agilent Technologies, Santa Clara, CA, USA).
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