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Description
DNase ⅠProduct Specification 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, 25C) Stability & Storage Store at 25 ~ 15 for 2 years Reference [1] Vanecko S, Laskowski M. Studies of the Specificity of Deoxyribonuclease I[J].
Product Specification
| 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 the Specificity of Deoxyribonuclease I[J]. Journal of Biological Chemistry, 1961, 236(236):3312-6. [2] Kienzle N, Young D, Zehntner S, et al. DNaseI treatment is a prerequisite for the amplification of cDNA from episomal-based genes[J]. Biotechniques, 1996, 20(4):612-6.
[3] Michael,R, Green, et
al. Human β-globin pre-mRNA synthesized in vitro is accurately spliced in
xenopus oocyte nuclei[J].Cell, 1983, 32(3):681-694.
|
Background
DNase I
(Deoxyribonuclease I), can digest single or double-stranded DNA to produce mono
deoxynucleotides or single or double-stranded oligo deoxynucleotides, its
optimal 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 can
randomly 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 flat
ends or 1-2 nucleotides protruding.
Components
Storage Solution: 2 U/ul DnaseⅠ、10mM Tris-Hcl、2mM CaCl2、50%Glycerol (pH7.6, 25℃)
10*Reaction
Buffer: 100mM Tris-Hcl、25mM MgCl2、5mM CaCl2 (pH7.6, 25℃)
Protocol
This step is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled up according to experimental needs.
1）Add the following components in sequence
| 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.
Guidelines
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
Unit Definition
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