Glucose Limitation Carboy

• Make 20L stock 10X MDS:

• Fill plastic carboy with approx. 17L MilliQ H₂O
• Add with constant stirring:

• CaCl₂-2H₂O                20g
• NaCl 20g
• MgSO₄-7 H₂O                        100g
• KH₂PO₄ (monobasic) 200g
• (NH₄)₂SO₄                  1000g

o Bring up to 20L with MilliQ H₂O

o Aliquot into 2L media bottles

o Autoclave

       For one carboy:

• Autoclave carboy
• Measure ~7L Milli-Q water into a "measuring" carboy
  

• Add the following to ~900ml in a 1L graduated cylinder and mix well:

• 1000x vitamins 10mL
• 1000x metals 10mL
• Glucose                                      8g

• Add contents of cylinder to "measuring" carboy
• Measure out 1L of 10X MDS  into the cylinder and  add to the "measuring" carboy
• Rinse cylinder with Milli-Q water into carboy
• Adjust volume to 10L

• Mix for  ~5min  with magnetic strirrer
Filter sterilize into autoclaved carboy

Dunham, Maitreya. http://genomics.princeton.edu/dunham/MDchemostat.pdf

Modified  by  S. Silverman  6/2006