In biochemical experiments, the buffer recipe is often the top priority of the experiment, and a truly reliable recipe can definitely change the fate of the experiment.

MOPS, 3-(N-Morpholino)propanesulfonic acid, is a biological buffer, the buffer range is 6.5~7.9, which is suitable for the study of electron transfer and phosphorylation in chloroplast thin layer; it can be prepared into a variety of agar, which is used as a non-toxic buffer in the culture of Streptomyces coelicolus and the production of cephalosporins in the limit culture medium. It can be used as the electrolyte system component in isoelectric focusing electrophoresis (IEF) of two-dimensional gel electrophoresis. It can also be used in Northern hybridization as a buffer for RNA separation and transfer.

Such an important buffer, what is the recipe? This article summarizes the recipe for almost all MOPS buffers for your reference.

（1）MOPS Buffer (10×)

Reagent	Quantity (for 1 L)	Final concentration
MOPS free acid, ultrapure	41.86 g (or 46.26 g of the sodium salt)	0.2 M
Sodium acetate anhydrous	4.1 g (or 6.8 g of acetate • 3H2O)	0.05 M
Na2EDTA (0.5M, pH 7.5)	20 mL	0.01 M

Bring the pH to 7.0 with NaOH (or if the MOPS sodium salt is used, then use glacial acetic acid to adjust the pH). Do not autoclave. Store in the dark, covered with foil, at room temperature. The solution will turn yellow with time, but this is normal.

（2）MOPS buffer (5X)

Reagent	Quantity (for 100 mL)	Final concentration (5X)
MgSO4	0.1204 g	10 mM
MOPS	10.463 g	0.5 M
NaCl	14.61 g	2.5 M

Dissolve the reagents listed above in 50 mL of H2O. Adjust the pH to 7.5 with NaOH, and then add H2O to 100 mL. Filter to sterilize, and store in the dark.

（3）MOPS salts

Reagent	MOPS	Tricine	FeSO4•7H2O	NH4Cl	K2SO4	CaCl2•2H2O	MgCl2	NaCl
Quantity	400mM	40mM	0.1mM	95mM	2.8mM	5μM	5.3mM	0.5M

Dissolve in a final volume of 1 liter, and sterilize by filtration. Add 10 μl of micronutrients per liter before use.

（4）MOPS-EGTA buffer

Reagent	Quantity (for 1 L)	Final concentration
MOPS	20.9 g	0.1 M
MgSO4	0.24 g	2 mM
EGTA	0.38 g	1 mM
NaCl	29.2 g	0.5 M

Dissolve the reagents above in 800 mL of diethyl pyrocarbonate (DEPC)-treated H2O. Adjust the pH to 7.5 with NaOH and then add DEPC-treated H2O to give a final volume of 1 L. Sterilize the buffer using a 0.22-μm filter. The buffer can be stored at room temperature for several months.

（5）MOPS electrophoresis buffer

0.2 M MOPS (pH 7.0), 20 mM sodium acetate, 10 mM EDTA (pH 8.0).

For 10x solution: Dissolve 41.8 g of MOPS in 700 ml of sterile Diethyl pyrocarbonate (DEPC)-treated H2O. Adjust the pH to 7.0 with 2 N NaOH. Add 20 ml of Diethyl pyrocarbonate (DEPC)-treated 1 M sodium acetate and 20 ml of Diethyl pyrocarbonate (DEPC)-treated 0.5 M EDTA (pH 8.0). Adjust the volume of the solution to 1 liter with Diethyl pyrocarbonate (DEPC)-treated H2O. Sterilize the solution by passing it through a 0.45-μm Millipore filter, and store it at room temperature protected from light. The buffer yellows with age if it is exposed to light or is autoclaved. Straw-colored buffer works well, but darker buffer does not.

（6）PFA-MOPS-EGTA fixation buffer

Reagent	Quantity (for 50 mL)	Final concentration
Paraformaldehyde	2 g	4% (w/v)
NaOH	(1 N) 5 mL	/
HCl	(1 N) 5 mL	/
MOPS-EGTA buffer	to 50 mL	/

Make fresh. To dissolve paraformaldehyde, add 2 g to 5 mL of 1 NaOH. Heat to 60°C to dissolve, and add 5 mL of 1 N HCl and 35 mL of MOPS-EGTA buffer. Adjust the pH to 7.5 if necessary and then add MOPS-EGTA buffer to give a final volume of 50 mL. Store the buffer at 4°C for several days.

Hope that these recipes could help you success in your experiments.

Edited by Suzhou Yacoo Science Co., Ltd.

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