| Species | Bovine Pancreatic |
| Synonyms | DNASE,Deoxyribonuclease-1 |
| Expression System | E.coli |
| Molecular Weight | 72kDa (Reducing) |
| Purity | >95% by SDS-PAGE&HPLC |
| Conjugation | Unconjugated |
| Tag | His Tag, MBP Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 10 mM Tris-HCl, 2 mM CaCl2 ,50% Glycerol，(pH 7.6, 25°C) |
| Stability & Storage | Store at -25 ~ -15℃for 2 years |
| Reference |
[1] Vanecko S, Laskowski M. Studies of theSpecificity of Deoxyribonuclease I[J]. Journal of Biological Chemistry, 1961,236(236):3312-6. [2] Kienzle N, Young D, Zehntner S, et al. DNaseItreatment is a prerequisite for the amplification of cDNA from episomal-basedgenes[J]. Biotechniques, 1996, 20(4):612-6. [3] Michael,R, Green, etal. Human β-globin pre-mRNA synthesized in vitro is accurately spliced inxenopus oocyte nuclei[J].Cell, 1983, 32(3):681-694. |
DNase I(Deoxyribonuclease I), can digest single or double-stranded DNA to produce monodeoxynucleotides or single or double-stranded oligo deoxynucleotides, itsoptimal working pH range is 7-8. DNase I activity is dependent on Ca2+and can be activated by other bivalent metal ions such as Mg2+, Mn2+,Zn2+, etc. In the presence of Mg2+, the enzyme canrandomly recognize and cut any site on any strand of DNA. In the presence of Mn2+,two strands of DNA can be cut at the same site to form sticky ends with flatends or 1-2 nucleotides protruding.
StorageSolution: 2 U/ul DnaseⅠ、10mM Tris-Hcl、2mM CaCl2、50%Glycerol (pH7.6, 25℃)
10*ReactionBuffer: 100mM Tris-Hcl、25mM MgCl2、5mM CaCl2 (pH7.6, 25℃)
This step is suitablefor linearization of 1 μg DNA (≥100 nt) and can be scaled up according toexperimental needs.
1）Add the following components insequence
| Components | Volume |
Plasmid DNA |
1μg DNA |
10*Reaction Buffer |
2μl |
DnaseⅠ (2U/μl) |
1μl |
RNase-free ddH2O |
Up to 50μl |
2）Incubate at 37°C 1 h.
1. EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation 2. Please avoid repeated freeze-thaw cycles
The results of 1μg pBR322 plasmid digestion separated under different quantity of DnaseⅠ, The reaction was incubated for 10 minutes at 37°C, and 1% agarose gel was used for electrophoresis analysis after reaction.
M, marker;
Lane 1 1μg pBR322;
Lane 2 1μg pBR322 add 4U DNase I
Lane 3 1μg pBR322 add 2U DNase I
Lane 4 1μg pBR322 add 1U DNase I
Lane 5 1μg pBR322 add 0.5U DNase I
Lane 6 1μg pBR322 add 0.25U DNase I
Lane
7 1μg pBR322 add 0.125U DNase I
1μg (R: reducing condition, N: non-reducing condition).
