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Team:Cambridge/Protocols/CCMB80
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| - | + | =CCMB80 buffer Recipe= | |
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| + | This buffer is used when making chemically competent cells | ||
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* 10 mM KOAc pH 7.0 (10 ml of a 1M stock/L or 0.98g/L) | * 10 mM KOAc pH 7.0 (10 ml of a 1M stock/L or 0.98g/L) | ||
* 80 mM CaCl<sub>2</sub>.2H<sub>2</sub>O (11.8 g/L) | * 80 mM CaCl<sub>2</sub>.2H<sub>2</sub>O (11.8 g/L) | ||
Revision as of 18:20, 23 August 2012
CCMB80 buffer Recipe
This buffer is used when making chemically competent cells
- 10 mM KOAc pH 7.0 (10 ml of a 1M stock/L or 0.98g/L)
- 80 mM CaCl2.2H2O (11.8 g/L)
- 20 mM MnCl2.4H2O (4.0 g/L)
- 10 mM MgCl2.6H2O (2.0 g/L or 5ml of 2M stock/L)
- 10% glycerol (100 ml/L)
- Fill up with DI water AFTER regulating pH
- adjust pH DOWN to 6.4 with 0.1N HCl if necessary
- adjusting pH up will precipitate manganese dioxide from Mn containing solutions.
- sterile filter and store at 4°C
- slight dark precipitate appears not to affect its function
