DNase I Solution (1 unit/μL), RNase-free

Thermo Scientific DNase I removes unwanted DNA from cell lysates to improve protein extraction efficiency.

Features of RNase-free DNase I:

• Degrades and removes unwanted DNA from samples
• Cleaves both single-stranded and double-stranded DNA
• Compatible with Thermo Scientific Pierce Cell Lysis Reagents
• Reduces viscosity of bacterial lysates (protein extracts) to facilitate pipetting

Deoxyribonuclease I (DNase I) is a single, glycosylated polypeptide that degrades unwanted single- and double-stranded DNA. The enzyme works by cleaving DNA into 5' phosphodinucleotide and small oligonucleotide fragments. 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. RNase-free DNase is useful for any application requiring the digestion of DNA in which it is crucial to avoid damage to RNA.

General information about the use of DNase I:

• Calcium ions are required for activity of DNase I. Trace amounts of Ca++ may be present at high enough concentration for DNase I to be active, however use of EGTA or calcium-free buffers can reduce DNase I activity to undetectable levels.
• High levels (i.e., 100 mM) of monovalent ions such as Na+ and K+ will decrease DNase I activity
• DNase I is inactivated by heating to 65°C for 10 minutes
• Kunitz unit: 1 Kunitz unit is the amount of enzyme required to cause an increase of 0.001 A260nm/min/mL at 25°C in 0.1M NaOAc, pH 5.0 due to degradation of highly polymerized DNA
• Degradation assay units: 1 unit is defined as the amount of enzyme required to completely degrade 1 μg of plasmid DNA in the 10 minutes at 37°C in 10 mM Tris·HCl, pH 7.5, 50 mM MgCl₂, 13 mM CaCl₂.

Specifications:

• Quantity: 1000 units (1 mL)
• Concentration: 1 unit/μL±20%
• Unit definition: 1 unit completely degrades 1 μg of plasmid DNA in 10 minutes at 37°C. One DNase I unit is equivalent to 0.3 Kunitz units.
• RNase Contamination: No ribonuclease activity detectable based on incubation with RNA transcript.
• Source: E. coli containing cloned gene encoding bovine DNase I
• Molecular Weight: Approx. 29,000
• Formulation: Bovine DNase I in 50 mM Tris-HCl (pH 7.5), 10 mM CaCl₂, 50% glycerol
• Supplied with: 1 mL of 10X Reaction Buffer 100 mM Tris-HCl (pH 7.5), 25 mM MgCl₂, 1 mM CaCl₂, 50 mM EDTA

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