News

All categories

Home page

The formula and precautions of MOPS buffer as a phosphorus-free medium

Release time:

2021-06-22

In many previous articles, we have introduced some configuration methods of MOPS medium, but did not talk about the specific items that need to be paid attention to during the configuration process. Today this article will list some items in great detail. For those who have not yet configured Friends who have used MOPS medium, we have prepared some configuration plans for MOPS medium. Hope to give you some guidance and help.

MOPS Minimal Medium

1. Mix the above ingredients and adjust the pH to 7.2 with about 300 microliters of 10M NaOH.

2. Filter and disinfect. It can be stored at 4 degrees for up to 1 month.

3. Before use, add 10 ml of 100X carbon source (when appropriate-we usually use a final concentration of 0.1% glucose).

10X MOPS mix

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

Ingredients	Firmware	Gram
MOPS	209.3	83.72
Tricine	179.2	7.17

2. Add 10 M KOH to make the final pH value 7.4 (10 to 20 ml)

3. Bring the total volume to 440ml.

4. Prepare a fresh FeSO4 solution and add it to the MOPS / Tricine solution:

Ingredients	Firmware	g	H2O volume (ml)	Stock concentration (M)
FeSO4 •7H2O	278	0.028	10	0.01

5. Add the following solutions to MOPS/tricine/FeSO4 (please refer to the methods below to make each method): mix in the order shown!

Composition	volume
1.9M NH 4 chlorine	50ml
0.276M K2SO4	10ml
0.02M calcium chloride 2•2H2O	0.25ml
2.5 M Magnesium Chloride 2	2.1 ml
5 M Sodium Chloride	100ml
Micronutrient stock	0.2ml
Autoclaved milliQ H2O	387ml
Full	1000ml

6. Filter and sterilize with a 0.2 micron filter with a capacity of 1L

7. Aliquot into 100 or 200ml sterile plastic bottles and freeze at -20°.

10 times the method used in the MOPS mix

Make separately and mix the specified amount into the specified volume. Store at room temperature.

Ingredients	Firmware	Stock Concentration (M)	Gram	Volume (ml)	Notes
NH4Cl	53.49	1.9	50.82	500
Potassium 2SO4	174.3	0.276	4.8	100
Calcium chloride 2•2H2O	147	0.02	0.294	100
Magnesium chloride 2	203.3	2.5	50.75	100
Sodium chloride	58.44	5	292.2	1000

Micronutrient stock solution (100ml)

Mix everything in 40ml of autoclaved milliQ H2O to bring the total volume to 50ml. Store at room temperature.

Ingredients	Molecular formula	Firmware	g 50ml
Ammonium molybdate	（NH4）6Mo7O24•4H2O	1235.9	0.009
Boric acid	h3BO3	61.83	0.062
Cobalt chloride	CoCl2	237.9	0.018
Copper sulfate	CuSO4	249.7	0.006
Manganese chloride	MnCl2	197.9	0.040
Zinc sulfate	ZnSO4	287.5	0.007