2× NPB
| Amount of reagent | Final concentration |
|---|---|
| 10 mL sucrose (e.g., Sigma-Aldrich SX1075) from a 1.5 m solution that has been filter-sterilized and stored at −20°C | 500 mm |
| 900 μL HEPES (e.g., Sigma-Aldrich H3375) from a 1 m, pH 7.7 solution that has been filter-sterilized and stored at −20°C | 30 mm |
| 120 μL EDTA (e.g., J.T. Baker 8991-01) from a 0.5 m, pH 8.0 solution | 2 mm |
| 300 μL spermidine trihydrochloride (e.g., Sigma-Aldrich S2501) from a 100 mm solution that has been aliquoted and stored at −20°C | 1 mm |
| 120 μL spermine tetrahydrochloride (e.g., Sigma-Aldrich S1141) from a 100 mm solution that has been aliquoted and stored at −20°C | 0.4 mm |
| 60 μL DL-dithiothreitol (DTT; e.g., Sigma-Aldrich D9779) from a 1 m solution that has been aliquoted and stored at −20°C | 2 mm |
| 18.5 mL sterile Nanopure H2O |
Freshly prepare 30 mL. Adjust pH to 7.7 with KOH. Store on ice.
