Nutrient broth

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-pH: 7.0 ± 0.2 at 25 °C. -Incubation: 24 h at 35 °C aerobically.

Protocol tips
-pH: 7.0 ± 0.2 at 25 °C. -Incubation: 24 h at 35 °C aerobically.

Publication protocol

In order to prepare SCFB, bacterial species i.e., Bacillus cereus (Bc), Clostridium tetani (Ct),
Listeria monocytogenes (Lm), and Neisseria meningitides (Nm) were grown separately in 100 mL of
Nutrient broth (NB) and human serum supplemented RPMI 1640 media (10 mL) for 24 h at 37 ◦C.
Anaerobic conditions were maintained by providing 5% CO2. After incubation, each of the cultural
broth was centrifuged at 5000 rpm for 10 min; the pellet was discarded and each supernatant was
separately air-dried overnight at ambient temperature. Dried broth was scrapped off by adding 2 mL
of D/W. Each SCFB was filter-sterilized using a 0.2 micron membrane Millipore filter. SCFBs were
marked as Bc, Ct, Lm, and Nm. Blank broths of both of the above-mentioned media were air-dried
and filter-sterilized likewise, which served as the control.

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Manufacturer protocol

Download the product protocol from Merck Millipore for Nutrient broth below.

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