• Up to 50X more activity and 350% greater catalytic efficiency than conventional DNase I
  
  
• Efficiently degrades DNA in solutions containing up to 0.25 M salt, unlike wild-type DNase I
  
  
• Efficiently digests DNA to oligonucleotides
  
  
• Vastly superior in clearing DNA templates from in vitro transcription reactions
  
  
• RNase-free and recombinant in origin: Purified from a source that is 10⁷-fold lower in RNase activity than bovine pancreas
  
  

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DNase I is an indispensable tool for removing DNA from molecular biology reactions. This enzyme cleaves double-stranded DNA non-specifically to leave 5' phosphorylated oligodeoxynucleotides. As a result, DNase I is commonly used to clear DNA contamination from RNA samples prior to RT-PCR. Yet DNase I is not without its limitations. Conventional DNase I has a poor affinity for DNA and cleaves DNA of low concentration very inefficiently. In addition, DNase I is very salt-sensitive; as little as 20 mM NaCl can reduce the activity of the enzyme by 30%. Thus, the enzyme is not very effective in reactions that require even modest amounts of salt, such as in vitro transcription reactions. Finally, DNase I is purified from bovine pancreas, one of the richest natural sources of RNase A. The threat of contaminating RNase activity in DNase I preparations requires that the enzyme be exhaustively purified. In spite of these limitations, the DNase I that researchers use today is the very same enzyme that was first characterized by Kunitz more than a half-century ago.

TURBO DNase: A New Hyperactive DNase with Superior Properties to Wild-type DNase I
TURBO DNase was developed using a protein engineering approach that introduced amino acid changes into the DNA binding pocket of wild-type DNase I. These changes markedly increase the affinity of the protein for DNA. The result is a versatile enzyme that has a 6-fold lower K_m for DNA, and an ability to maintain at least 50% of peak activity in solutions approaching 200 mM monovalent salt, even when the DNA concentration is in the nanomolar (nM) range. The flexibility of TURBO DNase activity is particularly evident when comparing the efficiency of DNA removal from in vitro transcription buffer, as shown in Figure 1. In this case, TURBO DNase reduces the input 50 kb lambda DNA to digested fragments that are roughly 100 times smaller than those treated with conventional DNase I. This qualitative result can be more precisely quantified using real-time RT-PCR (Table 2). When in vitro transcription reactions are treated with either DNase I or TURBO DNase, TURBO DNase removes 63X more of the input plasmid DNA tempate than the wild-type enzyme. As a result, TURBO DNase is the enzyme of choice for eliminating plasmid DNA templates from transcription reactions, particularly for the synthesis of RNA standards used in RT-PCR and generating RNA for microinjection and transfection experiments.

The proficiency of TURBO DNase in binding very low concentrations of DNA means that the enzyme is particularly effective in removing trace quantities of DNA contamination. This becomes important for complete removal of DNA from a sample, since the cleavable DNA substrate is reduced as the DNase reaction proceeds. TURBO DNase thus has a functional advantage over wild-type DNase due to its superior affinity for DNA. This is best exploited in RT-PCR applications, where even a few copies of DNA can lead to a false positive outcome by PCR. As shown in Table 1, TURBO DNase is more effective than wild-type DNase in reducing DNA detection by PCR.

View additional data showing the advantages of TURBO DNase over wild-type DNase I. Read a more detailed discussion of why TURBO DNase is superior to wild-type DNase I .

TURBO DNase is provided in two convenient sizes, 1000 U and 5000 U. A 10X Reaction Buffer is provided with the enzyme.

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