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 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, makes a good buffering agent for many biological systems requiring neutral pH. [[HEPES]] is a chemically similar pH buffering compound. | ||
==Recipe for 10x MOPS buffer== | ==Recipe for 10x MOPS buffer, 1 L== | ||
[[Image:800px-MOPS.png|right|200px|thumb|chemical structure of MOPS, 3-(N-morpholino) propanesulfonic acid]] | |||
* 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) | ||
* | |||
* 3.7 g EDTA, | |||
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* filter sterilise or autoclave | * filter sterilise or autoclave | ||
* store at room temperature | * store at room temperature | ||
* protect from light; do not use if the solution | * protect from light; do not use if the solution is dark (yellow is ok) | ||
==Final concentration | ==Final concentration in 10x stock== | ||
* 400 mM MOPS (buffering) | * 400 mM MOPS (buffering) | ||
* 100 mM NaAc | * 100 mM NaAc | ||
* 10 mM [[EDTA]] (nuclease inhibition by Mg2+ chelation) | * 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== | ==Stability of MOPS== | ||
Contrary to common belief, MOPS is sufficiently heat-stable to be autoclaved. | 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 [http://books.google.de/books?id=h5lGFPz054oC&pg=PA201&dq=farrel+rna+mops&lr=&ei=RAGsSb_8NouYMp7enJIF&hl=en]. Straw coloured buffer is good but do not use darker buffer. | ||
==Some OWW protocols which use MOPS== | ==Some OWW protocols which use MOPS== | ||
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== External links == | == External links == | ||
* [http://en.wikipedia.org/wiki/MOPS Wikipedia MOPS] | * [http://en.wikipedia.org/wiki/MOPS Wikipedia MOPS] | ||
* [http://www.fermentas.com/techinfo/electrophoresis/rnaloading.htm#MOPS_ MOPS recipe at Fermentas] | |||
* [http://www.sigmaaldrich.com/catalog/ProductDetail.do?N4=M5755|SIGMA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC premade MOPS 10x from Sigma] | |||
* [http://cshprotocols.cshlp.org/cgi/content/full/2006/3/pdb.rec8332?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=mops&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT&text_only=true CSH Protocols MOPS] [[Image:Padlock-closed.png]] | |||
[[Category:Material]] and [[Category:Buffers]] | |||
Latest revision as of 02:58, 10 January 2012
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, makes a good buffering agent for many biological systems requiring neutral pH. HEPES is a chemically similar pH buffering compound.
Recipe for 10x MOPS buffer, 1 L
- 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 is dark (yellow is ok)
Final concentration 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 good 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
