MOPS Minimal Medium

Updated 5/5/03. Now this medium is available commercially.

Updated 6/14/02. Our protocol for preparing Neidhardt MOPS Minimal Medium.

Reference: F. C. Neidhardt, P. L. Bloch, and D. F. Smith. 1974. Culture medium for enterobacteria. J Bacteriol 119(3): 736-747 [PubMed Central].

MOPS Minimal Medium

10X MOPS mixture

100 ml

 

0.132 M K2HPO4

10 ml

 

milliQ H2O

880 ml

 

1mg/ml thiamine

0.1 ml

(optional - we do not use thiamine because it does not affect the growth rate of E. coli K-12 MG1655 in this medium.)

TOTAL

990 ml

 

1. Mix ingredients above and adjust the pH to 7.2 with approximately 300 microliters 10 M NaOH.
2. Filter sterilize. Can be stored at 4 degrees for up to 1 month.
3. Before use add 10 ml 100X carbon source (as appropriate - we typically use a final concentration of 0.1% glucose).

10X MOPS Mixture

1. In a 1 L beaker with a stir bar, add the following to ~300 ml milliQ H2O:

Component FW grams

MOPS

209.3

83.72

Tricine

179.2

7.17

2. Add 10 M KOH to a final pH of 7.4 (10 to 20 ml)
3. Bring total volume to 440 ml
4. Make fresh FeSO4 solution and add it to the MOPS/Tricine solution:

Component FW grams H2O vol (ml) stock conc (M)
FeSO4•7H2O

278

0.028

10 0.01

5. Add the following solutions to the MOPS/tricine/FeSO4 solution (see below how to make each of these):

Mix in the order shown!

Component Volume

1.9 M NH4Cl

50 ml

0.276 M K2SO4

10 ml

0.02 M CaCl2•2H2O

0.25 ml

2.5 M MgCl2

2.1 ml

5 M NaCl

100 ml

Micronutrient stock

0.2 ml

Autoclaved milliQ H2O

387 ml

TOTAL

1000 ml

6. Filter sterilize with 1 L capacity 0.2 micron filter
7. Aliquot into sterile 100 or 200 ml plastic bottles and freeze at -20°.

Stocks used in 10X MOPS mixture

Make each separately, mixing the amount indicated into the specified volume. Store at room temp.

Component FW stock conc (M) grams vol (ml) note

NH4Cl

53.49

1.9

50.82

500

 

K2SO4

174.3

0.276

4.8

100

 

CaCl2•2H2O

147

0.02

0.294

100

 

MgCl2

203.3

2.5

50.75

100

 

NaCl

58.44

5

292.2

1000

 

Micronutrient stock (100 ml)

Mix everything together in 40 ml autoclaved milliQ H2O, bring up total volume to 50 ml. Store at room temp.

Component Formula FW Grams for 50 ml

ammonium molybdate

(NH4)6Mo7O24•4H2O

1235.9

0.009

boric acid

H3BO3

61.83

0.062

cobalt chloride

CoCl2

237.9

0.018

cupric sulfate

CuSO4

249.7

0.006

manganese chloride

MnCl2

197.9

0.040

zinc sulfate

ZnSO4

287.5

0.007

Potasium phosphate K2HPO4 Solution:

NOTE!! Make sure you use dibasic K2HPO4, and do NOT use monobasic KH2PO4 !!!!! Can be stored at room temperature after autoclaving.

Component FW stock conc (M) grams Vol (ml) note
K2HPO4

173.2

0.132

23.0 1000 autoclave


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