In the process of large-scale mRNA production, the transcription template needs to be removed after transcription. DNase I can randomly decompose single-stranded or double-stranded DNA to the same degree to generate oligonucleotides with 5'-P ends. Under Mg2+ conditions, DNase I can cut double-stranded DNA at will.
DNase I (Deoxyribonuclease I, deoxyribonuclease I) was originally isolated from bovine pancreas, with a molecular weight of about 39 kD. Has a 5'-P terminal oligonucleotide. DNase I hydrolyzes single-stranded or double-stranded DNA, its activity is very dependent on the level of Ca2+, and can be activated by Mg2+ or Mn2+.
This product is a GMP-grade recombinant DNase I expressed by large-scale fermentation of Pichia pastoris. It is produced with medicinal specifications raw materials and strictly controlled host protein residues, nucleic acid residues, etc., and conforms to GMP standard product production and quality management procedures to ensure the production process And all raw and auxiliary materials can be traced back.
Quality requirements
| Project |
Standard |
Method |
| Exterior |
Clear liquid |
Visual inspection |
| Visible foreign body |
Compliance |
Chinese Pharmacopoeia 2020 Edition Fourth Part 1 Lamp Inspection Method (General Rule 0904) |
| pH value |
7.0-8.0 |
Chinese Pharmacopoeia 2020 Edition Part IV pH Determination Method (General Principle 0631) |
| active |
1.8KUml-2.2KU/ml |
004 Plasmid DNA degradation method |
| Purity |
≥95% |
Chinese Pharmacopoeia 2020 Edition Part IV High Performance Liquid Chromatography (General Principle 0512) |
| RNase residue |
Degradation of 293-RNA does not exceed 10% |
2U enzyme and 293-RNA. Incubate at 37°C for 1h |
| Bacterial endotoxin content |
≤10 EU/mg |
Chinese Pharmacopoeia 2020 Edition Fourth Gel Limit Test Method (General Rule 1143) |
| Exogenous DNA residue |
≤100 pg/mg |
Fluorescence quantitative PCR |
| Host protein residue |
≤50 ppm |
Chinese Pharmacopoeia 2020 Edition Part IV Method for the Determination of Bacterial Protein Residues (General Rule 3414) |
| Mycoplasma detection |
Feminine |
Mycoplasma detection kit |
| Heavy metal residue |
≤10 ppm |
Chinese Pharmacopoeia 2020 Edition Fourth Heavy Metal Inspection Method (General Principle 0821) |
|
Follow the following specifications for production
1. ISO 9001:2015, certified facility.
2. "GMP Appendix-Cell Therapy Products" State Drug Administration.
3. "General Introduction to Human Gene Therapy-Chinese Pharmacopoeia 2020" National Pharmacopoeia Commission.
4. USP Chapter <1043>, Ancillary Materials for Cell, Gene, and Tissue-Engineered Products are used as excipients in cell therapy, gene therapy and tissue engineering products.
5. USP Chapter <92>, Growth Factors and Cytokines Used in Cell Therapy Manufacturing Cytokines and growth factors used in the production of cell therapy products.
6.Ph. Eur. General Chapter 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.
Product Usage
1. Prepare an RNA sample without DNA.
2. In the RNA sample before the RT-PCR reaction, remove possible DNA contamination such as genomic DNA.
3. In vitro T7, T3, SP6 and other RNA polymerases catalyze the removal of DNA template after in vitro transcription.
4. Used for Footprinting (Footprinting) to analyze DNA-protein interactions.
5. Used with DNA Polymerase I for nick translation.
6. In the presence of divalent manganese ions, the DNA is fragmented to generate a library of random DNA fragments.
7. In the apoptosis TUNEL test, part of the genomic DNA is sheared as a positive control.
Preservation system
10 mM Tris-HCl (pH7.6); 2 mM CaCl2; 50% (v/v) Glycerol.
Precautions
1. Optimal pH: 7.0-8.0
2. Activator: DNase I requires divalent cations to achieve maximum activity.
3. Inhibitors: EDTA, EGTA, SDS.
4. Specificity: double-strand specific endonuclease that degrades DNA.
5. Put the enzyme on ice during use, and store it at -20°C after use.
G2327352