- Производитель:
- Roche
Enzyme Commission (EC) Number: | 3.1.21.1 ( BRENDA | IUBMB ) |
Properties: Double-strand-specific endonuclease that requires bivalent cations for maximal activity.
Activity: 10,000U/bottle according to Kunitz (+25°C; DNA as substrate).
Unit definition: One unit is the enzyme activity that causes an increase in the absorbance of 0.001 per minutes under assay conditions.
- DNase I, recombinant, grade I is used for:Eliminating DNA during protein isolation procedures
- Analysis of chromatin structure
- Eliminating DNA during sample preparation
Lyophilizate
Assay mixture: Calf thymus DNA (100 µg) is incubated with 20 to 50 U DNase I at +25 °C.
The increase in absorbance is measured at 260 nm.
Information Note: These conditions are used for the determination of activity,according to Kunitz, since the Kunitz assay results in high reproducibility and sensitivity. When using this enzyme for normal experimental purposes, Roche recommends using incubation buffers that are appropriate for a given application, e.g., as mentioned in the standard literature (Sambrook et al., Curr. Prot. Mol. Biol., etc.).
Storage conditions (working solution): Long-term Storage of the Dissolved Enzyme
The solvent generally recommended for DNase I is water. When DNase I is reconstituted in water, the solution can be kept for 2 days at 2 to 8 °C and for 1 month at -15 to -20 °C. For best results, prepare appropriate aliquots and avoid repeated freezing and thawing. However, for long-term storage, carefully dissolve DNase I in one of the buffers listed below. In buffer 1, the enzyme will be stable for several weeks. The enzyme will be stable for at least 18 months if it is dissolved in buffer 2, 3, or 4 and stored at the recommended temperatures.
Information note: Do not vortex the enzyme while dissolving!
Buffer 1: 20 mM Tris-HCl, 20 mM MgCl2, 5 mM CaCl2, 0.1 mM dithioerythritol, 0.1 mM EDTA, 50% (v/v) glycerol, pH 8. Store this solution at -15 to-25 °C; it will not freeze at this temperature and will be stable for several weeks.
Buffer 2: 20 mM Tris-HCl, 50 mM NaCl, 1 mM dithioerythritol, 100 µg/mL BSA, 50% glycerol (v/v), pH 7.6. Store this solution at -15 to -25 °C for up to 18 months; it will not freeze at this temperature.
Buffer 3: 20 mM Tris-HCl, 1 mM MgCl2, 50% (w/v) glycerol, pH 7.5. Store this solution at -15 to -25 °C for up to 18 months; it will not freeze at this temperature.
Buffer 4: 20 mM Tris-HCl, 1 mM MgCl2, pH 7.5. Aliquot in appropriate amounts (e.g., 10 µl), freeze the aliquots quickly on dry ice, and store at -60 °C or below for up to 18 months. Thaw only the amount needed for each experiment. Do not refreeze. Discard any leftover thawed solution.
Quality Level | 100 |
form | lyophilized |
packaging | pkg of 2 x 10,000 U |
Торговая марка | Roche |
shipped in | wet ice |
storage temp. | 2-8°C |
pictograms | GHS08 |
signalword | Danger |
hcodes | H317 - H334 |
pcodes | P261 - P280 - P284 - P304 + P340 - P333 + P313 - P342 + P311 |
RIDADR | NONH for all modes of transport |
WGK Germany | WGK 1 |
Flash Point F | does not flash |
Flash Point C | does not flash |
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