IGEM:Virginia 2012/Protocols/SSM Media Preparation

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Contents

Materials

BASAL MEDIUM to make 1 Liter 2 Liters 4 Liters 5 Liters
Glutamic acid, monosodium salt, 1H2O 10.72g 21.44g 42.88g 53.60g
Trisma Base 1.525g 3.05g 6.10g 7.63g
NaCl 2.50g 5.00g 10.00g 12.50g
KH2PO4 0.50g 1.00g 2.00g 2.50g
KCl *200 mg/mL 0.20g 0.40g 0.80g 1.00g
MgCl2 6H2O *100mg/mL (0.5 mM) 100mg 200mg 400mg 500mg
CaCl2 2H2O *20 mg/mL 20mg 40mg 80mg 100mg
Q.S. with deionized water to** 980mL 1960mL 3920mL 4900mL
  • Note: *Make stock solutions and add this amount.
  • Note: **These Volumes are to allow for 20 mL/L worth of supplements to be added later.
  • Note: For SSM-S, add 2.0 g/L of 2-ketoglutarate and pyruvate before pH'ing.
  • Note: if pouring blood plates, bring volume up to 100 mL/L LESS than above.
  • Note: it is convenient to make up the last three salts in a concentrated stock solution, such that you can use 1mL/liter of SSM.

Procedure

  • Add 2.9g of NaCl to a 1 L bottle.
  • Add in 25 mL of 7.5 pH Tris-Cl
  • Bring this to a volume of 500 mL by adding distilled water.
  • Autoclave (large liquid).

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!


References

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