RNase H Buffer (5×)
| Reagent | Quantity (for 10 mL) |
Final concentration (5×) |
|---|---|---|
| Tris–HCl (1 m, pH 7.5) | 1 mL | 100 mm |
| KCl (2 m) | 2.5 mL | 500 mm |
| MgCl2 (1 m) | 0.5 mL | 50 mm |
| DTT (1 m) | 0.5 mL | 50 mm |
| Sucrose | 2.5 g | 25% (w/v) |
| H2O | to 10 mL | |
| If this buffer is used infrequently, prepare a stock without DTT, aliquot into 1-mL aliquots, and add 50 μL of 1 m DTT to each aliquot before use. Store working aliquot with DTT for up to 1 mo at –20°C. | ||
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