Enzymes for Molecular Biology: Enzymes for Molecular Biology

RNase-Free DNase I (bovine pancreas) is an endonuclease useful in removing DNA that might interfere with the characterisation, manipulation, or use of RNA, or for any application requiring highly purified DNase I. It efficiently hydrolyses double- and single-stranded DNA to a mixture of short oligo- and mononucleotides.

Figure 1. DNA removal from in vitro transcription reactions using RNase-Free DNase I. A linearized DNA template was transcribed using T7 RNA polymerase according to standard invitro transcription conditions. Lane 1, kb ladder; Lane 2, DNA control; Lane3, transcription mixture; Lane 4, transcription mixture treated with 1MBU of RNase-Free DNase I for 15 minutes at 37°C.

Unit Definition

Using the standard 50µl assay volume, one Molecular Biology Unit (MBU) digests 1µg of pUC79 DNA to oligodeoxynucleotides in 10 minutes at 37°C in 33 mM Tris-acetate, pH 7.8, 66mM potassium acetate, 10mM magnesium acetate, and 0.5mM DTT 2mM CaCl2.

Storage Buffer

50% glycerol containing 10mM Tris-HCl, pH 7.5, 10mM CaCl2, and 10mM MgCl2.

Quality Control

No degradation of 1µg of a synthetic RNA transcript is detected by agarose gel electrophoresis following incubation with 10U of RNase-Free DNase I at 37°C for 1 hour.

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Protocols for: RNase-Free DNase I

RNase-Free DNase I Protocol

(catalogue number D9902K / D9905K / D9910K)

Please note: all protocols off site are the responsibility of the products supplier

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References

1. Kienzle, N. et al. (1996) BioTechniques 20, 612.
2. Sambrook, J. et al. (1989) in: Molecular Cloning: A Laboratory Manual (2nd ed.) Cold Spring Harbor Laboratory Press, New York.
3. Galas, D.J. and Schmitz, A. (1978) Nucleic Acids Res. 5, 3157.

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Applications

• Elimination of template DNA following in vitro synthesis of RNA with T7, SP6, or T3 RNA polymerase.
• Labelling of DNA by nick translation, in combination with Klenow or other DNA polymerases.
• Treatment of RNA prior to RT-PCR.¹
• Characterization of DNA-protein interactions by DNase I footprinting.^2,3

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