The M. lusitanicus double auxotrophic strain MS12 (leuA− and pyrG−), a derivative of the strain CBS 277.49 [31 (link)], was used in the present study. For spore collection, the strain was grown for 7 days on malt extract agar (MEA; 1% glucose, 0.25% yeast extract, 5% malt extract, 2% agar; pH 5.4) plates at 25 °C; then, the spores were washed with PBS by gentle scraping, and the spore suspensions obtained were maintained at 4 °C until use. For aerobic growth, 30 mL liquid minimal medium (YNB; 1% glucose, 0.15% ammonium sulphate, 0.15% sodium glutamate, 0.05% yeast nitrogen base without amino acids (BD Difco, Becton Dickinson, Franklin Lakes, NJ, USA), supplemented with 0.05% leucine and 0.05% uracil; pH 6.8) was inoculated to obtain a final concentration of 104 sporangiospores/mL. The cultures were kept at 25 °C for 24 h with constant shaking (i.e., 190 rpm). For the mycelium–yeast transition, the 24 h old culture grown aerobically was transferred into a 50 mL flask into 50 mL fresh YNB medium and placed for 4 h into a BBL GasPak Anaerobic System (Becton Dickinson, Franklin Lakes, NJ, USA) at 25 °C.
Bbl gaspak anaerobic system
Manufactured by BD
Sourced in United States
The BBL GasPak Anaerobic System is a laboratory equipment designed for the cultivation and growth of anaerobic microorganisms. It provides a controlled anaerobic environment to support the metabolic requirements of these organisms. The system includes components that generate and maintain an anaerobic atmosphere within an enclosed chamber.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bbl gaspak anaerobic system
1
Cultivation of M. lusitanicus Auxotrophic Strain
2
Isolation of Lactobacillaceae from Ostrich Feces
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lactobacillaceae members were isolated as per protocols described by Tsuchida et al. (2014) with a few modifications. One gram of feces from each ostrich was diluted in 9 mL of sterilized saline and mixed thoroughly. The slurry was further diluted to 10−2 with sterilized saline. A portion (100 µL) of the diluted samples was spread onto an LBS agar plate (Becton, Dickinson and Company, Sparks, MD, USA). The plates were then transferred into the BBL GasPak Anaerobic System (Becton, Dickinson and Company) and incubated anaerobically for 6 days at 39°C. Colonies on the agar plate were selected and inoculated into 2 mL of MRS broth (Becton, Dickinson and Company). The culture was then incubated for 4 d at 39°C. A full loop (10 µL) of the culture fluid was again streaked onto LBS agar and incubated anaerobically in a GasPak anaerobic system at 39°C. Finally, a single colony was selected, inoculated into 10 mL of MRS broth, and incubated for 4 d at 39°C.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!