Master Mix (4×)
| Reagent | Final concentration |
|---|---|
| TES–Tris (pH 7.0) | 200 mm |
| KCl | 400 mm |
| MgCl2 · 6H2O | 28 mm |
| Ethylene glycol tetraacetic acid (EGTA; pH 7.0) | 4 mm |
| TES is buffered with saturated Tris (tris(hydroxymethyl)aminomethane). The 4× master mix should be freshly prepared, but the separate stock solutions can be pre-prepared and stored at 4°C or frozen at −20°C for several months. The pH of the EGTA stock solution should be accurately determined at pH 7.0 using NaOH. We recommend freezing aliquots of EGTA for later use. | |
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