How do you store DNase?

Store up to 18 months at −15 to −25°C. The solution will not freeze. 20 mM Tris-HCl, 1 mM MgCl2, pH 7.5. Freeze in aliquots of approximately 10 µl quickly on dry ice; store up to 18 months at −60°C or below.

What is in DNase buffer?

The recipe of DNase buffer is: 100 mM Tris-HCl (pH 7.5), 25 mM MgCl2, 1 mM CaCl2. You can make it in DEPC treated water to get rid of RNase.

What does DNase do in our lysis buffer?

DNase I is commonly added to cell lysis reagents to remove the viscosity caused by the DNA content in bacterial cell lysates or to remove the DNA templates from RNAs produced by in vitro transcription. This grade of DNase is sufficient for protein work.

Does DNase I need magnesium?

DNase I requires activation through divalent metals; maximum activation is achieved with magnesium and calcium but manganese, cobalt, and zinc may also be used. If random cleavage or nicking is desired, magnesium ions should be used as an activator.

How long is DNase stable for?

The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room tempera- ture. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature.

How do you make DNase stock?

DNase I made up to 40,000 U/ml in storage buffer (-20°C freezer door) = stock solution. Dilute stock solution 1:40 in 10x reaction buffer = 1,000 U/ml = 1 U/ul = working dilution. Use at a 1:10 dilution: 1μl per 9 μl solution to be treated. Incubate at room temperature for 2hr.

How do you make a DNase solution?

Prepare 1M EDTA disodium salt dihydrate. Weigh out 3.72 g EDTA and place in 5 ml molecular biology grade water. Add a few NaOH pellets to get to pH 9 and dissolve EDTA (can warm to 45°C with stirring to aid in dissolution).

How do you make DNase RNase free?

Clean equipment with a detergent solution, rinse thoroughly with water and rinse with 95% ethanol to dry. Soak the equipment in a 3% hydrogen peroxide (H2O2) solution for ten minutes at room temperature. Rinse the equipment thoroughly with DEPC-treated water.

How do you remove DNA from a protein sample?

Phenol chloroform extraction, normally followed by ethanol precipitation, is the traditional method to remove protein from a DNA sample. In this procedure, the DNA solution is mixed with phenol and chloroform.

What is the role of DNase I?

DNASE 1. Deoxyribonuclease I (DNase I, encoded by DNASE1) is a specific endonuclease facilitating chromatin breakdown during apoptosis. DNase I activity is important to prevent immune stimulation, and reduced activity may result in an increased risk for production of antinucleosome antibodies, a hallmark of SLE.

How long is DNase stable at room temperature?

Storage Conditions and Product Stability The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room tempera- ture. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months.

How stable is DNase at room temperature?

The DNase Max Kit is stable at room temperature (15–25°C) for up to 6 months or at 2–8°C for 2 years with no loss of activity. Room temperature storage eliminates the need to aliquot and freeze stocks of DNase I enzyme and removes concerns about decreased enzyme activity due to freeze–thaw cycles.