Cleave and degrade unwanted single- and double-stranded DNA.
Thermo Scientific DNase I removes unwanted DNA from cell lysates to improve protein extraction efficiency.
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.
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
Available in two formats:
RNase-free (Part No. 89836), with vial of 10X Reaction Buffer
Protein extraction grade (Part No. 90083)
Thermo Scientific Lysozyme improves protein extraction efficiency. To demonstrate the benefit of using lysozyme and DNase I to process bacterial extracts, the extractions of two different-sized, over-expressed proteins were compared. Cell pellets from 50mL culture of E. coli BL-21 over-expressing green fluorescent protein (GFP) or GST-Ral Binding Protein (GST-RalBP) were lysed using B-PER Reagent with and with out lysozyme and DNase I. The soluble fractions were separated from the pellets, and the two fractions were analyzed by SDS-PAGE. Although GFP (32 kDa) was extracted equally well in the absence or presence of enzyme, GST-RalBP (75 kDa) required lysozyme and DNase I to be efficiently solubilized.
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., 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 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 1µg of plasmid DNA in the 10 minutes at 37°C in 10mM Tris•HCl, pH 7.5, 50mM MgCl2, 13mM CaCl2.
Specifications (Part No. 89836), RNase-Free:
Quantity: 1000 units (1mL)
Concentration: 1 unit/µL ± 20%
Unit definition: 1 unit completely degrades 1µg of plasmid DNA in 10 minutes at 37°C. One DNase I unit is equivalent to 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 50mM Tris-HCl (pH 7.5), 10mM CaCl2, 50% glycerol