DNase I GMP-grade
DNase I enzyme for DNA digestion in RNA workflows.
Product overview
DNase I (Deoxyribonuclease I) is a deoxyribonuclease that can efficiently digest both single-stranded and double-stranded DNA fragments. The activity of DNase I is highly dependent on Ca2+ concentration, and can be activated by Mg2+ or Mn2+. In the presence of Mg2+, DNase I cleaves DNA fragments randomly at any site; whereas in the presence of Mn2+, DNase I cleaves double-stranded DNA at approximately the same position on both strands, generating blunt-ended DNA fragments or sticky ends with 1-2 nucleotide overhangs. DNase I exhibits optimal endonuclease activity when the working pH ranges from 7 to 8, and it is widely applied to remove single-stranded and double-stranded DNA contaminants from various RNA samples.
Available versions
| Cat. no. | Product name | Formulation |
|---|---|---|
| GMP-BP-E04 | DNase I GMP-grade | |
| GMP-BP-E04-2K | DNase I GMP-grade | |
| GMP-BP-E04-20K | DNase I GMP-grade |
Features and applications
- Recombinantly expressed in Pichia yeast strain; activity: 2 U/μL.
- Unit definition: one unit (U) is defined as the amount of enzyme required to completely digest 1 μg of plasmid DNA in 10 min at 37°C.
- Quality control: protein purity ≥95%; free of exogenous RNase activity; residual host-cell DNA ≤100 pg/mg; residual host-cell protein <50 ppm; bacterial endotoxin <5 EU/mL.
- Preparation of DNA-free RNA samples for downstream RNA analysis.
- Digestion of linearized plasmid template following in vitro transcription.
- Removal of residual genomic DNA prior to RT-PCR.
- DNase I footprinting analysis of DNA-protein interactions.
Protocol
Notes
- Mix the reaction mixture by gentle pipetting; avoid vigorous vortexing.
- Keep the enzyme on ice throughout the procedure.
- Mix thoroughly before use. Aliquot for storage to avoid repeated freeze-thaw cycles.
- The optimal amount of DNase I GMP-grade should be optimized according to the specific experimental requirements.
DNA removal from RNA samples
- Using RNase-free microcentrifuge tubes or PCR tubes, prepare the reaction mixture as follows:
Components Volume 10×DNase I Reaction Buffer 1 μL DNase I GMP-grade (2 U/μL) 1 μL RNA Up to 1 μg RNase-free ddH2O To 10 μL - Mix gently and incubate at 37°C for approximately 30 min.
- To inactivate the enzyme, add EDTA to a final concentration of 5 mM and incubate at 65°C for 10 min.
Components
| Components | GMP-BP-E04-200 | GMP-BP-E04-2K | GMP-BP-E04-20K |
|---|---|---|---|
| 200U | 2KU | 20KU | |
| DNase I GMP-grade (2 U/μL) | 0.1 mL | 1 mL | 10 mL |
| 10×DNase I Reaction Buffer | 0.2 mL | 2 mL | 20 mL |
Performance data
1 Performance Comparison Test
Description: Digest 1 μg plasmid, perform gradient dilution of DNase I from different suppliers, and conduct electrophoresis to determine the activity of the DNase I from different suppliers.
Conclusion: During the process of removing the template after IVT, 1 μg of the plasmid was digested. Biori DNase I achieved complete digestion at a lower dosage than the DNase I from Supplier A and Supplier B.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | Store at -20±5°C. |
| Shipping | Ship frozen on dry ice or with validated frozen cold-chain packaging that maintains the specified frozen storage condition. |
| Handling | Avoid repeated freeze-thaw cycles; mix thoroughly after thawing where applicable. |
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