MOPS: Difference between revisions
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[[Image:800px-Mops2.JPG|right|200px|thumb|The other mops]] | [[Image:800px-Mops2.JPG|right|200px|thumb|The other mops [http://en.wikipedia.org/wiki/Pug] ]] | ||
'''MOPS''' is the common name for the buffering compound in MOPS buffer. MOPS stands for ''3-(N-morpholino) propanesulfonic acid'' and with a pKa of 7.20, MOPS is an good buffer for many biological systems at almost neutral pH. [[HEPES]] is a chemically similar pH buffering compound. | '''MOPS''' is the common name for the buffering compound in MOPS buffer. MOPS stands for ''3-(N-morpholino) propanesulfonic acid'' and with a pKa of 7.20, MOPS is an good buffer for many biological systems at almost neutral pH. [[HEPES]] is a chemically similar pH buffering compound. | ||
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* 83.7 g MOPS; MW 209.3 g/mol | * 83.7 g MOPS; MW 209.3 g/mol | ||
* 13.6 g Sodium Acetate, trihydrate; MW 136.1 g/mol (watch out: NaAc, anhydrous is only 82 g/mol or 8.2g) | * 13.6 g Sodium Acetate, trihydrate; MW 136.1 g/mol (watch out: NaAc, anhydrous is only 82 g/mol or 8.2g) | ||
* | * 3.7 g EDTA, disodium dihydrate; MW 372.24 g/mol (again check MW of the salt you stock) | ||
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* 200 mM MOPS - 41.9 g in 1 L | * 200 mM MOPS - 41.9 g in 1 L | ||
* 20 mM NaAc - 2.7 g | * 20 mM NaAc - 2.7 g | ||
* 10 mM [[EDTA]] - | * 10 mM [[EDTA]] - 3.7 g | ||
==Stability of MOPS== | ==Stability of MOPS== |
Revision as of 11:20, 2 March 2009
MOPS is the common name for the buffering compound in MOPS buffer. MOPS stands for 3-(N-morpholino) propanesulfonic acid and with a pKa of 7.20, MOPS is an good buffer for many biological systems at almost neutral pH. HEPES is a chemically similar pH buffering compound.
Recipe for 10x MOPS buffer
- 83.7 g MOPS; MW 209.3 g/mol
- 13.6 g Sodium Acetate, trihydrate; MW 136.1 g/mol (watch out: NaAc, anhydrous is only 82 g/mol or 8.2g)
- 3.7 g EDTA, disodium dihydrate; MW 372.24 g/mol (again check MW of the salt you stock)
- add 800 ml of nuclease free distilled water; mix to dissolve
- adjust to pH 7 with NaOH (prepared in nuclease free distilled water)
- fill to the final volume of 1000 ml
- filter sterilise or autoclave
- store at room temperature
- protect from light; do not use if the solution appears yellow
Final concentration of active compounds in 10x stock
- 400 mM MOPS (buffering)
- 100 mM NaAc
- 10 mM EDTA (nuclease inhibition by Mg2+ chelation)
Variation
Some protocols use a less concentrated MOPS buffer
- 200 mM MOPS - 41.9 g in 1 L
- 20 mM NaAc - 2.7 g
- 10 mM EDTA - 3.7 g
Stability of MOPS
Contrary to common belief, MOPS is sufficiently heat-stable to be autoclaved. Solution will turn yellow but this does not interfere with its buffering capacity. See, for example, Farrell RNA methods, p201 [2]. Straw coloured buffer is but do not use darker buffer.
Some OWW protocols which use MOPS
- Jacobs:Protocol RNA Agarose Gel
- Knight:NuPAGE electrophoresis
- Sauer:bis-Tris SDS-PAGE, the very best