Bacterial cell culture media Staphylococcus aureus

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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BD Tryptic Soy Agar

analitik kimya

Upstream tips	Protocol tips	Downstream tips
	Staphylococcus aureus USA300 LAC strain, a clinical isolate [14, 15], and Enterococcus faecalis OG1RF, a human isolate [16], were adopted as recipients for plasmid transfer experiments. All strains except E. faecalis OG1RF were primarily cultured in tryptic soy agar (TSA; BD Diagnostic Systems, Sparks, MD, USA) and tryptic soy broth (TSB; BD Diagnostic Systems) at 30 °C for 24 h to maintain their phenotypic traits. E. faecalis OG1RF traits were maintained in De Man-Rogosa-Sharpe (MRS; BD Diagnostic Systems) media at 30 °C for 24 h.

Protocol tips
Staphylococcus aureus USA300 LAC strain, a clinical isolate [14, 15], and Enterococcus faecalis OG1RF, a human isolate [16], were adopted as recipients for plasmid transfer experiments. All strains except E. faecalis OG1RF were primarily cultured in tryptic soy agar (TSA; BD Diagnostic Systems, Sparks, MD, USA) and tryptic soy broth (TSB; BD Diagnostic Systems) at 30 °C for 24 h to maintain their phenotypic traits. E. faecalis OG1RF traits were maintained in De Man-Rogosa-Sharpe (MRS; BD Diagnostic Systems) media at 30 °C for 24 h.

Protocol tips

Staphylococcus aureus USA300 LAC strain, a clinical isolate [14, 15], and Enterococcus faecalis OG1RF, a human isolate [16], were adopted as recipients for plasmid transfer experiments. All strains except E. faecalis OG1RF were primarily cultured in tryptic soy agar (TSA; BD Diagnostic Systems, Sparks, MD, USA) and tryptic soy broth (TSB; BD Diagnostic Systems) at 30 °C for 24 h to maintain their phenotypic traits. E. faecalis OG1RF traits were maintained in De Man-Rogosa-Sharpe (MRS; BD Diagnostic Systems) media at 30 °C for 24 h.

Manufacturer protocol Publication protocol 1 Relevant paper

BRAIN HEART INFUSION BROTH

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	-pH 7.4 ± 0.2 @ 25°C. -A satisfactory medium for blood culture can be prepared by adding 1g of agar per litre of Brain Heart Infusion Broth. Ensure that the agar is uniformly distributed in the sterile broth before dispensing into bottles.	-Tubes of Oxoid Brain Heart Infusion Broth which are not used the same day as sterilized should be placed in a boiling water bath for several minutes to remove absorbed oxygen, and cooled rapidly without shaking, just before use.

Protocol tips
-pH 7.4 ± 0.2 @ 25°C. -A satisfactory medium for blood culture can be prepared by adding 1g of agar per litre of Brain Heart Infusion Broth. Ensure that the agar is uniformly distributed in the sterile broth before dispensing into bottles.
Downstream tips
-Tubes of Oxoid Brain Heart Infusion Broth which are not used the same day as sterilized should be placed in a boiling water bath for several minutes to remove absorbed oxygen, and cooled rapidly without shaking, just before use.

Manufacturer protocol Publication protocol 1 Relevant paper

Nutrient agar

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.	-Final pH 6.8 +/- 0.2 at 25°C. - If desired, the medium can be enriched with 5-10% v/v sterile defibrinated blood.

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
-Final pH 6.8 +/- 0.2 at 25°C. - If desired, the medium can be enriched with 5-10% v/v sterile defibrinated blood.

Manufacturer protocol Publication protocol 1 Relevant paper

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