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AT Minimal Medium

AT Minimal Medium

Jun 18, 2021

Reference:  Tempe, 1977, PNAS 74:2848

AT is prepared as 20X buffer and 20X salts solutions that are supplemented with a carbon source.

20X AT Buffer

g/mol

g/L

Final Concentration (1X)

20X AT Buffer

g/mol

g/L

Final Concentration (1X)

KH2PO4

136.1

214

0.079 M

pH to 7.0 w/ NaOH

 

35 (solids)

0.044 M

20X AT Salts

g/mol

g/L

Final Concentration (1X)

20X AT Salts

g/mol

g/L

Final Concentration (1X)

(NH4)2SO4

132.14

40

0.015 M

MgSO4-7H2O (anhydrous,1.6)

246.5

3.2

0.6 mM

CaCl2-2H2O (anhydrous, 0.151)

147.0

0.2

0.06 mM

FeSO4-7H2O

278.0

0.1

0.027 mM

MnSO4-H2O

169

0.024

0.0071 mM

1X AT Medium

g/mol

g/L

Final Concentration (1X)

1X AT Medium

g/mol

g/L

Final Concentration (1X)

20X buffer

 

50mL/L

1X

20X salts

 

50mL/L

1X

50% glucose or 40% succinate

 

10mL/L

0.5%

+/- Bacto-agar

 

15

1.5%

  1. Prepare sterile, autoclaved stocks of 20X salts, 20X buffer and the carbon source. Add the appropriate amounts to either sterile deionized water (AT broth) or molten (55 oC) sterile 1.5% agar-water (AT agar); mix gently, but thoroughly.

  2. Iron will precipitate after autoclaving salts to form red iron oxides. (i.e., rust)

  3. After 1X broth made, there is no need to autoclave. So the process of 1X preparation should be sterile.   DI should be autoclaved separately.

  4. The sugar (e.g., 50% glucose, 40% succinate) needs to be filtered before storage.