Applications
- 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
- Characterization of DNA-protein interactions by DNase I footprinting.2,3
RNase-Free DNase I (bovine pancreas) is an endonuclease useful in removing
DNA that might interfere with the characterization, manipulation, or
use of RNA, or for any application requiring highly purified DNase I,
such as nick translation. It efficiently hydrolyzes dsDNA and ssDNA into
a mixture of short oligonucleotides and mononucleotides.
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.
References
- 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.
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