MUELLER-HINTON BROTH

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Oxoid Mueller-Hinton Broth will require supplementation with the divalent cations Mg++ and Ca++ after sterilisation2. The CLSI recommend the following cation levels Ca++, 20-25mg/litre; Mg++, 10-12.5mg/litre. -pH 7.3 ± 0.1 @ 25°C

Protocol tips
-Oxoid Mueller-Hinton Broth will require supplementation with the divalent cations Mg++ and Ca++ after sterilisation2. The CLSI recommend the following cation levels Ca++, 20-25mg/litre; Mg++, 10-12.5mg/litre. -pH 7.3 ± 0.1 @ 25°C

Publication protocol

Growth curves of the strains DSM 10303, DSM 13414, and field isolates Bb5/12 and Bb24/12 were generated over 48 hours by measuring the optical density OD600 on a UV-visible spectrophotometer (Varian Cary-100, Mulgrave Victoria, Australia), as well as by culture-based enumeration. For all four isolates an overnight culture was carried out in CAMHB (Oxoid, Wesel, Germany) at 35°C in ambient air. The overnight culture was adjusted to 0.5 McFarland standard and a 1:100 dilution was carried out in 0.9% saline. From this diluted bacterial suspension 100 μl was added to 100 ml of each medium to reach a start cfu/mL of approximately 103 cfu/mL.
Isolates were cultured in cation-adjusted Mueller-Hinton broth (CAMHB), CAMHB plus 2% (v/v) lysed horse blood, Brain-Heart-Infusion (BHI) (Becton Dickinson, Heidelberg, Germany) and Caso broth (Sigma-Aldrich, Seelze, Germany), which are frequently used media for the cultivation of Bordetella spp.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for MUELLER-HINTON BROTH below.

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