Bacterial cell culture media Streptococcus pyogenes

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

Protocol tips
-GAS is grown on agar plates produced from Todd-Hewitt broth supplemented with 0.2% yeast extract (THY). All steps should be performed using sterile technique. GAS will remain viable on plates for only 5–7 days after streaking if stored at room temperature. GAS does not survive at 4°C.Sterile wood sticks or inoculating loops used.Optimal growth of GAS is seen in 5% CO2; however, GAS will also grow in ambient air albeit a little slow.
-Final pH: 7.8 ± 0.2 at 25°C.
Protocol tips
The experiments used the following human S. pyogenes strains: S. pyogenes ATCC ® 19615 ™, S. pyogenes ATCC ® 49399 ™ and a clinical isolate obtained from a patient with pharyngitis (Bacteriology, Queen Elizabeth II Hospital, Halifax, NS, Canada). The strains were routinely cultured in Brain Heart Infusion (BHI) (Oxoid, Thermo Fisher Scientific, Waltham, MA, USA) media at 37 °C under aerobic conditions.
BRAIN HEART INFUSION BROTH

Thermo Fisher Scientific

Upstream tips
-Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C.
Protocol tips
-Reaction of 3.7% w/v aqueous solution at 25°C. pH : 7.4±0.2.
-Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.
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