User:Jarle Pahr/Buffers: Difference between revisions
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! Buffer!! Composition !! Use !! Storage !! Hazardous chemicals? || Comments | ! Buffer!! Composition !! Use !! Storage !! Hazardous chemicals? || Comments | ||
|- | |||
|TAE||40mM Tris, 20mM acetic acid, and 1mM EDTA||Electrophoresis||Room temperature || || | |||
|- | |- | ||
| Qiagen EB|| 10 mM Tris-HCl, pH 8.5 || DNA elution and storage || Room temperature || - || | | Qiagen EB|| 10 mM Tris-HCl, pH 8.5 || DNA elution and storage || Room temperature || - || | ||
|- | |- | ||
|TE || 10 mM Tris-HCl, pH 8.5 + 10 mM Tris-HCl, pH 8 + 1 mM EDTA ||DNA elution and storage || ? || - ||EDTA inhbitis nucleases and also inhibits sequencing reactions. | |TE (Tris-EDTA) || 10 mM Tris-HCl, pH 8.5 + 10 mM Tris-HCl, pH 8 + 1 mM EDTA ||DNA elution and storage || ? || - ||EDTA inhbitis nucleases and also inhibits sequencing reactions. | ||
|- | |||
|NEBuffer 1 ||10mM Bis-Tris-Propane-HCl, 10mM MgCl2,1mM DTT,pH 7.0@25°C|| Restriction digestion||-20C|| - ||CIP shows 50 % activity in NEBuffer 1, compared with 100 % activity in NEBuffers 2-4. | |||
|- | |- | ||
|NEBuffer | |NEBuffer 2||50mM NaCl,10mM Tris-HCl,10mM MgCl2,1mM DTT,pH 7.9@25°C||Restriction digestion||-20C || - || | ||
|- | |- | ||
|NEBuffer | |NEBuffer 3||100mM NaCl,50mM Tris-HCl,10mM MgCl2,1mM DTT,pH 7.9@25°C||Restriction digestion||-20C || - || | ||
|- | |- | ||
|NEBuffer | |NEBuffer 4||50mM Potassium Acetate,20mM Tris-acetate,10mM Magnesium Acetate,1mM DTT,pH 7.9@25°C||Restriction digestion||-20C || - || | ||
|- | |- | ||
| | |Phosphate Buffered Saline (PBS)|| || ||8 g/L NaCl, 0.2 g/L KCl, 1.44 g/L Na2HPO4, 0.22 g/L KH2PO4. Adjust pH to 7.4 with HCl.||http://cshprotocols.cshlp.org/content/2006/1/pdb.rec8247.full?text_only=true | ||
|} | |} | ||
'''Transformation and Storage Solution (TSS):''' | |||
LB with 10 wt/vol % polyethylene glycol (PEG), 5 % v/v DMSO, 50 mM Mg2+, pH 6.5. | |||
HEPES: | |||
http://en.wikipedia.org/wiki/HEPES | |||
MOPS: | |||
http://en.wikipedia.org/wiki/MOPS |
Latest revision as of 05:47, 14 May 2013
Qiagen EB buffer: 10 mM Tris-HCl, pH 8.5.
See also http://people.mbi.ucla.edu/sumchan/qiagenbuffer.html
Buffer | Composition | Use | Storage | Hazardous chemicals? | Comments |
---|---|---|---|---|---|
TAE | 40mM Tris, 20mM acetic acid, and 1mM EDTA | Electrophoresis | Room temperature | ||
Qiagen EB | 10 mM Tris-HCl, pH 8.5 | DNA elution and storage | Room temperature | - | |
TE (Tris-EDTA) | 10 mM Tris-HCl, pH 8.5 + 10 mM Tris-HCl, pH 8 + 1 mM EDTA | DNA elution and storage | ? | - | EDTA inhbitis nucleases and also inhibits sequencing reactions. |
NEBuffer 1 | 10mM Bis-Tris-Propane-HCl, 10mM MgCl2,1mM DTT,pH 7.0@25°C | Restriction digestion | -20C | - | CIP shows 50 % activity in NEBuffer 1, compared with 100 % activity in NEBuffers 2-4. |
NEBuffer 2 | 50mM NaCl,10mM Tris-HCl,10mM MgCl2,1mM DTT,pH 7.9@25°C | Restriction digestion | -20C | - | |
NEBuffer 3 | 100mM NaCl,50mM Tris-HCl,10mM MgCl2,1mM DTT,pH 7.9@25°C | Restriction digestion | -20C | - | |
NEBuffer 4 | 50mM Potassium Acetate,20mM Tris-acetate,10mM Magnesium Acetate,1mM DTT,pH 7.9@25°C | Restriction digestion | -20C | - | |
Phosphate Buffered Saline (PBS) | 8 g/L NaCl, 0.2 g/L KCl, 1.44 g/L Na2HPO4, 0.22 g/L KH2PO4. Adjust pH to 7.4 with HCl. | http://cshprotocols.cshlp.org/content/2006/1/pdb.rec8247.full?text_only=true |
Transformation and Storage Solution (TSS):
LB with 10 wt/vol % polyethylene glycol (PEG), 5 % v/v DMSO, 50 mM Mg2+, pH 6.5.
HEPES:
http://en.wikipedia.org/wiki/HEPES
MOPS: