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

CSF samples were collected from the local diagnostic lab of Karachi with the consent and approval
of lab authority. All persons gave their informed consent prior to inclusion in the study. Samples were
collected aseptically and kept at −20 ◦C until further use. For the isolation of etiological agents, 10 µL
of each of the CSF sample was streaked separately on different culture media, i.e., nutrient agar, blood
agar, and chocolate agar. The experiment was run in duplicate for the isolation of both aerobic and
anaerobic bacteria. One set of the media plates were incubated aerobically at 37 ◦C. For the isolation of
the anaerobic etiological agents, another set of the above-mentioned media was incubated at 37 ◦C in
the presence of 5% CO2. All plates were initially incubated for 24 h; incubation was then continued up
to 48 h. Plates were first observed after 24 h and then after 48 h. Bacteria were identified on the basis of
visual colonial characteristics, cellular morphology, and standard biochemical reactions.

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Manufacturer protocol

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

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