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

Description

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. Two grades of DNAse are offered, one sufficient for protein work and one that is useful for any application requiring the digestion of DNA in which it is crucial to avoid damage to RNA.

Highlights:

• 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
• Supplied as 50% glycerol stocks in 10mM Tris-HCl pH 7.5, 10mM CaCl₂, 10mM MgCl₂
• Available in two convenient formats: RNase-tested to protect RNA (Part No. 89836) and convenient protein extraction grade (Part No. 90083)

• Calcium ions are required for DNase I activity, and trace amounts of Ca++ may be present at high enough concentrations for DNase I to be active; however, the use of EGTA or calcium-free buffers can reduce DNase I activity to undetectable levels
• High levels (i.e., 100mM) 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 in absorbance at 260nm of 0.001/min/mL at 25°C in 0.1 M NaOAc, pH 5.0, due to the degradation of highly polymerized DNA
• Degradation assay units: 1 unit is defined as the amount of enzyme required to completely degrade 1mg of plasmid DNA in 10 minutes at 37°C in 10mM Tris-HCl, pH 7.5, 50mM MgCl₂, 13mM CaCl₂

• Visual: Clear liquid
• Concentration: 1 Unit/mL
• Activity: ≥ 100,000 Units/mg protein
• RNase Contamination: 20 Units release < 2% non-ethanol precipitable when incubated with 3H-labeled Poly(A) RNA
• Functional Analysis: 1μL enzyme degrades < 5% RNA but degrades 100% DNA

• Visual: Clear, colorless liquid, free of insoluble material
• Units: ≥2500 units/mL
• Unit definition: 1 unit is defined as the amount of enzyme required to produce an increase in absorbance at 260nm of 0.001/min/mL at 25°C of highly polymerized DNA

• Elimination of viscosity caused by DNA in protein cell lysates
• Removal of DNA templates from RNAs produced by in vitro transcription