The wastewater was concentrated using the flocculation method using 3.0% w/v beef extract solution (Sigma-Aldrich, St. Louis, USA), pH 9.5 in 0.05 M glycine buffer. Each sewage water (SW1, SW2, and SW3) sample (500 mL) was acidified (Michael-Kordatou et al. 2020 (link)) at pH 3.5 ± 0.1 and 10 mL of the beef extract was agglomerated by the addition of 0.1 M HCl at pH 3.0 followed by stirring for 10 h. The stirred suspension was centrifuged at 10,000×g for 30 min at 4°C. The pellet was dissolved in 8 mL of phosphate buffered saline (PSB) and the RNA was extracted using NucliSENS® miniMAG® system (BioMerieux, Marcy l Etoile, France) according to the manufacturer’s instructions.
Beef extract solution
Manufactured by Merck Group
Sourced in United States
Beef extract solution is a laboratory reagent used as a nutrient source for the cultivation of microorganisms in microbiology applications. It provides essential nutrients, vitamins, and growth factors necessary for the growth and maintenance of various bacterial cultures.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using beef extract solution
1
Wastewater Viral RNA Extraction Protocol
2
Wastewater Virus Concentration and RNA Extraction
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The wastewater concentration was conducted by direct flocculation [45 (link)] using beef extract solution (3.0% w/v, pH 9.5; Sigma-Aldrich, St. Louis, USA) in glycine buffer (0.05 M). Briefly, 500 mL of the wastewater sample was acidified (pH 3.5 ± 0.1) and 10 mL of beef extract flocculated by the addition of 1 and 0.1 M HCl (pH 3.5 to 3.0; a visible floc formed) was added. The suspension was stirred for 10 h to allow the viruses present to adsorb to the flocs and subsequently centrifuged at 10,000× g for 30 min at 4 °C. The pellet was dissolved in 8 mL of phosphate-buffered saline (PBS) and RNA isolated with the NucliSENS® miniMAG® system (BioMérieux, Marcy l’Etoile, France) according to the manufacturer’s instructions.
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