Streptococcus mutans Growth

Generic Tips

• Strains can grow on Tryptic Soy Broth plates (TSB) or TSB + Blood plates. There is no hemolysis on blood plates.
• Strains are usually grown on Brain Heart Infusion (BHI).
• Strain s-763 take 2 days to grow on plates from the freezer; please plan accordingly! Other strains take ~24 hours to grow.
• SMUR is a low peptide liquid media used for transformation, BHI is used otherwise

• Shaking is not needed. Notice that these cells have a more granular appearance, as they form tight clusters of cells.
• Overnight cultures in BHI begin to die off; please do not plan on using them for more than the next day, if at all.
• Less death is evident in SMSDM (S. mutans Semi-defined Media); same for BHI plates; we notice few cells surviving after 48 hours.

SMSDM

	For 500ml (100% vitamins)	For 500ml (50% vitamins)	For 3ml (50% vitamins)
Base	447.5ml	449ml	2.7ml
Solution I — C+Y vitamins	3ml	1.5ml	9ul
Solution IB	2ml	2ml	12ul
Solution II — Metals	5ml	5ml	15ul
Solution III — Unstables	2.5ml	2.5ml	15ul
10% Glucose	40ml	40ml	240ul

Base Medium

Base Media 1.11x concentration

Compound	Amount	Final Concentration
K2HPO4	5.61g	58mM
KH2PO4	1.13g	15mM
(NH4)2SO4	734mg	10mM
NaCl	1.14g	35mM
Casamino Acids	1.10g	0.2%

• Fill to 450ml
• pH to 7.4