Recombinant DNase I (RNase-free), 10KU
Cat. No.: SB-G3342-10KU
Size: 10KU
Description
DNase I is an endonuclide enzyme, which can degrade single and double stranded DNA to 5 'phosphate group, 3' hydroxyl terminal monodeoxynucleotide or oligonucleotide. DNase I is calcium-dependent and can be activated by magnesium and divalent manganese ions. In the presence of magnesium ion, DNase I could randomly splice any site of double-stranded DNA. In the presence of divalent manganese ions, DNase I can splice DNA double strands at the same site, forming flat ends, or sticky ends with 1 or 2 nucleotides protruding.
Main Uses
- Preparing RNA samples without DNA;
- Genomic DNA residues were removed from RNA samples before RT-PCR reaction.
- In vitro T7, T3, SP6 and other RNA Polymerases catalyze the removal of DNA templates in the RNA post-transcription system;
- Nick translationin DNA markers;
- Generating libraries of random DNA fragments;
- Apoptosis Tunel detection of partial genomic DNA shear as positive control.
- Features: specifically degrades DNA, but cannot degrade RNA.
- Source: Recombinant expression of the pichia coli strain carrying the Bovine Pancreatic DNase I gene.
- Definition of enzyme activity: The amount of enzyme required for complete degradation of 1 µg pBR322 vector DNA was defined as one enzyme activity unit after incubation at 37℃ for 10 min.
- Inactivation or inhibition: DNase I can be fully inactivated by heating at 75℃ for 10 min.
- Purity: SDS-PAGE purity ≥95%, RNase free; 10 U/mu L.
Storage and Handling Conditions
Transport with wet ice; Store at -20℃, valid for 12 months.
Composition
| Component |
G3342-10KU |
| Recombinant DNase I (RNase-free) |
1 mL |
| 10×DNase I Reaction Buffer |
11×1 mL |
| 25 mM EDTA |
6×1 mL |
Assay Protocol / Procedures
Reference use (To prepare RNA samples without DNA as an example)
1. Reference reaction system:
| Component |
Volume |
| RNA |
1 μg |
| Recombinant DNase I (RNase-free) |
1 U |
| 10×DNase I Reaction Buffer |
1 μL |
| RNase-free Water |
To 10 μL |
2. Set the reaction system according to step 1, mix gently, then centrifugal the liquid to the bottom of the tube;
3. Incubate at 37℃ for 30 minutes, then add 0.5 μL 25 mM EDTA to the reaction system to stop the reaction;
4. DNase I was completely inactivated after incubation at 75℃ for 10min.
Note
1. When using the product, the enzyme should be stored in an ice box or ice bath and stored immediately after use at -20ºC.
2. For your safety and health, please wear lab coat and disposable gloves to operate.
