RNase-Free DNase I
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Includes 10X Reaction Buffer.
RNase-Free DNase I
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- Degrade dsDNA and ssDNA without damaging important RNA in your sample or reaction
- Digest away unwanted DNA without digesting RNA
- Use in a variety of applications such as in vitro transcription and RT-PCR
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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, such as nick translation. It efficiently hydrolyses dsDNA and ssDNA into a mixture of short oligonucleotides and mononucleotides.
- Elimination of template DNA following in vitro synthesis of RNA with T7, SP6, or T3 RNA polymerase.
- Labeling of DNA by nick translation, in combination with Klenow or other DNA polymerases.
- Treatment of RNA prior to RT-PCR. 1
- Characterisation of DNA-protein interactions by DNase I footprinting. 2,3
Figure 1. DNA removal from in vitro transcription reactions using RNase-Free DNase I. A linearised DNA template was transcribed using T7 RNA polymerase according to standard in vitro transcription conditions. Lane 1, kb ladder; Lane 2, DNA control; Lane 3, transcription mixture; Lane 4, transcription mixture treated with 1 MBU of RNase-Free DNase I for 15 minutes at 37 °C.
- Kienzle, N. et al. (1996) BioTechniques 20, 612.
- Sambrook, J. et al. (1989) in: Molecular Cloning: A Laboratory Manual (2nd ed.) Cold Spring Harbor Laboratory Press, New York.
- Galas, D.J. and Schmitz, A. (1978) Nucleic Acids Res. 5, 3157.
Unit Definition: One Molecular Biology Unit (MBU) of RNase-Free DNase I digests 1 µg of pUC19 DNA to oligodeoxynucleotides in 10 minutes at 37 °C under standard assay conditions.
Storage Buffer: 50% glycerol containing 10 mM Tris-HCl (pH 7.5), 10 mM CaCl2, and 10 mM MgCl2.
DNase I 10X Reaction Buffer: 100 mM Tris-HCl (pH 7.5), 25 mM MgCl2, and 5 mM CaCl2.
Quality Control: No degradation of 1 µg of a synthetic RNA transcript is detected by agarose gel electrophoresis following incubation with 10 U of RNase-Free DNase I at 37 °C for 1 hour.