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| Product Name | Recombinant DNase I (RNase-Free, E. coli-Expressed) |
| Catalog No. | IVDRM-0051 |
| Description | Recombinant DNase I (Deoxyribonuclease I) is an endonuclease that digests both single-stranded and double-stranded DNA. It hydrolyzes phosphodiester bonds, yielding mono- and oligodeoxynucleotides carrying 5'-phosphate and 3'-hydroxyl groups. The enzyme works optimally at pH 7–8, requires Ca2+ for activity, and can be activated by divalent metal ions such as Co2+, Mn2+, and Zn2+. In the presence of Mg2+, DNase I cleaves each strand of double-stranded DNA at random, independent sites; in the presence of Mn2+, it cleaves both strands at approximately the same site, generating blunt ends or sticky ends with 1–2 nucleotide overhangs. Produced recombinantly in E. coli with a His tag, this RNase-free preparation is suitable for the treatment of a wide variety of RNA samples. |
| Intended Use | Removal of contaminating DNA from RNA samples, including template DNA removal after in vitro transcription and rRNA removal during RNA-seq (NGS) library preparation. |
| Principle / Technology | DNase I hydrolyzes phosphodiester bonds in DNA, producing mono- and oligodeoxynucleotides with 5'-phosphate and 3'-OH termini. Its activity is Ca2+-dependent and is further activated by divalent cations such as Co2+, Mn2+, and Zn2+. With Mg2+, DNase I introduces random nicks on either strand of double-stranded DNA; with Mn2+, both strands are cut at essentially the same position, yielding blunt ends or ends with 1–2 nucleotide overhangs. |
| Sample Type | RNA samples; active on single-stranded DNA, double-stranded DNA, chromatin, and DNA-RNA hybrids as substrates. |
| Performance Range / Specifications | High digestion efficiency: rapid and efficient DNA degradation upon incubation at 37 °C. RNase-free: no detectable RNase residue. Substrate versatility: acts on single-stranded DNA, double-stranded DNA, chromatin, and DNA-RNA hybrids. |
| Specificity | Specifically degrades DNA with no effect on RNA. |
| Reaction Conditions / Protocol | Optimal working pH range is 7–8. Incubate at 37 °C for rapid, efficient DNA degradation. Activity depends on Ca2+ and is activated by divalent metal ions such as Co2+, Mn2+, and Zn2+; cleavage pattern differs with Mg2+ (random single-strand nicks) versus Mn2+ (concerted double-strand cuts). |
| Storage Conditions | Store at -25 to -15 °C. Aliquoting is recommended; avoid repeated freeze-thaw cycles. |
| Shelf Life | 2 years |
| Package Specifications | 1000 U / 5000 U / 5×1000 U |
| Quality Control | RNase residue testing: DNase I is incubated with substrate RNA and RNA band integrity is compared by agarose gel electrophoresis, confirming no detectable RNase contamination. In template DNA removal during in vitro transcription and rRNA removal during RNA library construction, performance is comparable to leading competing products. |
| Source / Origin | Recombinant DNase I expressed in E. coli; carries a His tag. |
| pH Range / Optimal pH | Optimal working pH range: 7–8 |
| Expiration Date / Stability | Stable for 2 years when stored at -25 to -15 °C; aliquot to avoid repeated freeze-thaw cycles. |
| Application Notes / Precautions | Aliquot upon receipt and avoid repeated freeze-thaw cycles. Suitable for RNA sample treatment, removal of DNA templates after in vitro transcription, and RNA-seq library preparation; yields are comparable to leading competing products. |
For research use only, not for clinical use.
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