Bacterial cell culture media Bordetella bronchiseptica

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.

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Found 3 matching solutions for this experiment

Brain heart infusion bhi agar 20 plates BD Difco BBL

Science Equip

Upstream tips	Protocol tips	Downstream tips
	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. [17–19]. Incubation was performed at 35°C in ambient air. Sampling was performed every two hours during the first 26 hours and, thereafter every six hours.

Protocol tips
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. [17–19]. Incubation was performed at 35°C in ambient air. Sampling was performed every two hours during the first 26 hours and, thereafter every six hours.

Protocol tips

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. [17–19]. Incubation was performed at 35°C in ambient air. Sampling was performed every two hours during the first 26 hours and, thereafter every six hours.

Manufacturer protocol Publication protocol 1 Relevant paper

CASO Broth

Sigma-Aldrich

Upstream tips	Protocol tips	Downstream tips
-Store prepared media below 8°C, protected from direct light. Store dehydrated powder, in a dry place, in tightly-sealed containers at 2-25°C.	-Anticoagulants such as sodium polyanetholesulfonate (Cat. No. P2008) or sodium citrate (Cat. No. 71635) may be added to the broth prior to sterilisation. 5 to 10 ml of blood may be added to 50 ml of medium.

Upstream tips
-Store prepared media below 8°C, protected from direct light. Store dehydrated powder, in a dry place, in tightly-sealed containers at 2-25°C.
Protocol tips
-Anticoagulants such as sodium polyanetholesulfonate (Cat. No. P2008) or sodium citrate (Cat. No. 71635) may be added to the broth prior to sterilisation. 5 to 10 ml of blood may be added to 50 ml of medium.

Manufacturer protocol Publication protocol 1 Relevant paper

MUELLER-HINTON BROTH

Thermo Fisher Scientific

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

Manufacturer protocol Publication protocol 1 Relevant paper

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