Brain Heart Infusion Broth for Microbiology

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	supplement the media with 44 mL/L glycerol, 30 mg/L hemin and 10 mg/L nicotinamide adenine dinucleotide (NAD)

Protocol tips
supplement the media with 44 mL/L glycerol, 30 mg/L hemin and 10 mg/L nicotinamide adenine dinucleotide (NAD)

Publication protocol

All reagents were from Sigma Aldrich, New South Wales, Australia unless otherwise stated. Glycerol stocks of S. pneumoniae D39 (ATCC#7466) and NTHi 289 [7] were streaked out onto blood agar or chocolate agar plates respectively for single colonies and incubated at 37°C. S. pneumoniae was incubated anaerobically using the BD GasPak™ EZ Anaerobic Pouch System [BD Diagnostics, Australia]. Following overnight incubation, three colonies were selected from each agar plate and used to inoculate culture media. S. pneumoniae was grown statically in brain heart infusion (BHI) broth at 37°C and NTHi was grown with shaking in BHI supplemented with 44 mL/L glycerol, 30 mg/L hemin and 10 mg/L nicotinamide adenine dinucleotide (NAD). Both strains were grown to mid-log phase (OD600nm 0.55 – 0.65), counted at 100 × magnification using a Helber bacteria counting chamber [ProSciTech, Queensland, Australia] and then either subjected to the different treatments (heat, ethanol or freezing) or used fresh on the day of preparation. All cultures were streaked onto agar plates and incubated at 37°C overnight to check purity. For cultures that were to be frozen, heat-inactivated FCS [SAFC Biosciences, New South Wales, Australia] was added to the mid-log phase culture to give a final concentration of 20% FCS and 1 mL aliquots were stored in cryovials at -80°C. FCS was the only cryoprotectant used for storage of these bacterial stocks.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Bacterial cell culture media Streptococcus pneumoniae using Brain Heart Infusion Broth for Microbiology from Sigma-Aldrich.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Brain Heart Infusion Broth for Microbiology below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Bacterial cell culture media Streptococcus pneumoniae using Brain Heart Infusion Broth for Microbiology from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.