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An Animal-Free DNase I

Safety of biological products is also an emerging issue. Eliminating animal tissues and animal-derived components from the production process prevents potential transmission of pathogens. Materials derived from bovine sources are of special concern, due to known transmission of bovine spongiform encephalopathy (BSE) to humans via ingestion of cow products. Regulatory agencies are increasingly restrictive in this regard, prompting the pharmaceutical industry to remove high risk raw materials from manufacturing. Ambion's rDNase I now addresses these issues by providing a recombinant replacement for pancreatic bovine DNase I.


rDNase I Performance
Ambion's rDNase I meets or exceeds the performance of native bovine DNase I. It digests DNA to oligonucleotide-sized products or shorter. Yet its reaction kinetics are identical to that of native DNase I. Thus, rDNase I can be directly substituted for native DNase I with equivalent units.

The recommended reaction conditions are the same as Ambion's standard DNase I. Incubate 1-2 U of rDNase I per 1 g DNA for 30 min at 37C in a buffer consisting of 10 mM Tris-HCl, pH 7.5, 2.5 mM MgCl2, 0.5 mM CaCl2.


Quantitative Removal of Contaminating DNA
To demonstrate the efficiency of rDNase I in removing residual DNA, we measured genomic DNA contamination by real-time PCR before and after DNase I treatment. A total of 0.5 g of human genomic DNA was incubated in 50 l, and digested with 2 U of DNase I or rDNase I in a volume of 50 l for 30 min at 37C. One-tenth of this reaction volume (equivalent to 50 ng genomic DNA) was added to a 25 l real-time PCR reaction. The eff
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NM522 contains an F episome, which is required for production of ssDNA and for blue/white color selection. To maintain the F, NM522 should be grown on minimal (M-9) medium....
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