Streptococcus mutans Growth: Difference between revisions
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PreDec2022>EricMiller (Created page with "* Strains can grow on Tryptic Soy Broth plates (TSB) or TSB + Blood plates. There is no hemolysis on blood plates. * Strains take 2 days to grow on plates from the freezer; p...") |
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= Generic Tips = | |||
* Strains can grow on Tryptic Soy Broth plates (TSB) or TSB + Blood plates. There is no hemolysis on blood plates. | * Strains can grow on Tryptic Soy Broth plates (TSB) or TSB + Blood plates. There is no hemolysis on blood plates. | ||
* Strains take 2 days to grow on plates from the freezer; please plan accordingly! | * 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 = | |||
{| class="wikitable" style="margin:auto" | |||
! !! 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 == | |||
{| class="wikitable" style="margin:auto" | |||
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 |
Latest revision as of 14:41, 15 May 2024
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 concentrationCompound | 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