DNase I脱氧核糖核酸酶I内切酶|Deoxyribonuclease I(DNase I) GMP-grade
产品说明书
FAQ
COA
已发表文献
DNase I is an endonuclease that can digest single-stranded and double-stranded DNA to produce single deoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides. It can hydrolyze the phosphodiester bond to produce monodeoxynucleotides and oligodeoxynucleotides containing 5'-phosphate groups and 3'-OH groups. The average digestion product is the smallest polytetranucleotide. DNase I can catalyze many forms of DNA, such as single-stranded DNA, double-stranded DNA, and even chromatin (its cutting rate is affected by histones).The optimum pH range is 7-8. The activity of DNase I depends on Ca2+ and can be activated by divalent metal ions, such as Co2+, Mn2+, Zn2+, etc. 5 mM Ca2+ can protect the enzyme from being hydrolyzed. In the presence of Mg2+, the enzyme can recognize and cut any site on any strand of DNA randomly; and in the presence of Mn2+, it can recognize two strands of DNA at the same time and cut at almost the same site to form blunt ends, Or sticky ends with 1-2 nucleotides protruding. DNase I is widely used in the preparation of DNA-free RNA; remove the template DNA after in vitro transcription; prepare DNA-free RNA before RT-PCR and RT-qPCR reactions; combine with DNA polymerase I to perform DNA labeling through nick translocation; DNA fragmentation library construction.
This product is produced in accordance with GMP process requirements, and the product is provided in liquid form.
Product Properties
|
Source |
Recombinant E. coli with DNase I gene |
|
Optimum Temperature |
37℃ |
|
Storage Buffer |
10 mM Tris-HCl pH 7.6，2 mM CaCl2，50%(v/v) Glycerol |
|
Unit Definition |
The amount of enzyme required to increase the absorbance at 260 nm of the reaction solution by 0.001 in 1 minute at 25°C and pH 5.0 using calf thymus DNA as the substrate is defined as one activity unit (Kunitz Unit).(The reaction buffer is: 10 mM Tris-HCl pH 7.6, 2.5 mM MgCl2, 0.5 mM CaCl2, 1 μg plasmid DNA.) |
Contents
|
Contents No. |
Name |
Catalog No./Specification |
||||
|
10611ES76 （500 U） |
10611ES84 （2,000 U） |
10611ES92 （10,000 U） |
10611ES98（100 KU） |
|||
|
10611 |
Deoxyribonuclease I (DNase I) GMP-grade (2 U/μL) |
250 μL |
1 mL |
5 mL |
50 mL |
|
Shipping and Storage
Deoxyribonuclease I (DNase I) GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.
Experimental methods
Plasmid template digestion
1. Reaction system:
Use the RNase-free centrifuge tube and pipette tip to prepare the following reaction system:
|
10× DNase I Buffer* |
1 μL |
|
DNase I |
1 μL |
|
RNA |
x |
|
Rnase-free ddH2O |
Up to 10 μL |
【Note】1×DNase I Buffer: 10 mM Tris-HCl, 2.5 mM MgCl2, 0.5 mM CaCl2, pH7.6 @25℃
2. Reaction conditions
37℃, 15-30 min later, add a final concentration of 2.5 mM EDTA solution and mix well at 65℃ for 10 min. The processed template can be used in subsequent reactions such as capping reaction
DNase Ⅰ inactivation or inhibition
After adding EDTA to a final concentration of 2.5 mM, heating at 65°C for 10 min can inactivate DNase I. Phenol and chloroform extraction can also inactivate DNase I. The following conditions all have significant inhibitory effect on DNase I: Metal ion chelating agents, zinc ions with a concentration of millimoles/liter, 0.1% SDS, DTT, mercaptoethanol and other reducing agents,the salt concentrations above 50-100 mM.
Note
1. Enzymes should be stored in an ice box or on an ice bath when used, and should be stored at -20°C immediately after use.
2. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.
HB230406
DNase I is an endonuclease that can digest single-stranded and double-stranded DNA to produce single deoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides. It can hydrolyze the phosphodiester bond to produce monodeoxynucleotides and oligodeoxynucleotides containing 5'-phosphate groups and 3'-OH groups. The average digestion product is the smallest polytetranucleotide. DNase I can catalyze many forms of DNA, such as single-stranded DNA, double-stranded DNA, and even chromatin (its cutting rate is affected by histones).The optimum pH range is 7-8. The activity of DNase I depends on Ca2+ and can be activated by divalent metal ions, such as Co2+, Mn2+, Zn2+, etc. 5 mM Ca2+ can protect the enzyme from being hydrolyzed. In the presence of Mg2+, the enzyme can recognize and cut any site on any strand of DNA randomly; and in the presence of Mn2+, it can recognize two strands of DNA at the same time and cut at almost the same site to form blunt ends, Or sticky ends with 1-2 nucleotides protruding. DNase I is widely used in the preparation of DNA-free RNA; remove the template DNA after in vitro transcription; prepare DNA-free RNA before RT-PCR and RT-qPCR reactions; combine with DNA polymerase I to perform DNA labeling through nick translocation; DNA fragmentation library construction.
This product is produced in accordance with GMP process requirements, and the product is provided in liquid form.
Product Properties
|
Source |
Recombinant E. coli with DNase I gene |
|
Optimum Temperature |
37℃ |
|
Storage Buffer |
10 mM Tris-HCl pH 7.6，2 mM CaCl2，50%(v/v) Glycerol |
|
Unit Definition |
The amount of enzyme required to increase the absorbance at 260 nm of the reaction solution by 0.001 in 1 minute at 25°C and pH 5.0 using calf thymus DNA as the substrate is defined as one activity unit (Kunitz Unit).(The reaction buffer is: 10 mM Tris-HCl pH 7.6, 2.5 mM MgCl2, 0.5 mM CaCl2, 1 μg plasmid DNA.) |
Contents
|
Contents No. |
Name |
Catalog No./Specification |
||||
|
10611ES76 （500 U） |
10611ES84 （2,000 U） |
10611ES92 （10,000 U） |
10611ES98（100 KU） |
|||
|
10611 |
Deoxyribonuclease I (DNase I) GMP-grade (2 U/μL) |
250 μL |
1 mL |
5 mL |
50 mL |
|
Shipping and Storage
Deoxyribonuclease I (DNase I) GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.
Experimental methods
Plasmid template digestion
1. Reaction system:
Use the RNase-free centrifuge tube and pipette tip to prepare the following reaction system:
|
10× DNase I Buffer* |
1 μL |
|
DNase I |
1 μL |
|
RNA |
x |
|
Rnase-free ddH2O |
Up to 10 μL |
【Note】1×DNase I Buffer: 10 mM Tris-HCl, 2.5 mM MgCl2, 0.5 mM CaCl2, pH7.6 @25℃
2. Reaction conditions
37℃, 15-30 min later, add a final concentration of 2.5 mM EDTA solution and mix well at 65℃ for 10 min. The processed template can be used in subsequent reactions such as capping reaction
DNase Ⅰ inactivation or inhibition
After adding EDTA to a final concentration of 2.5 mM, heating at 65°C for 10 min can inactivate DNase I. Phenol and chloroform extraction can also inactivate DNase I. The following conditions all have significant inhibitory effect on DNase I: Metal ion chelating agents, zinc ions with a concentration of millimoles/liter, 0.1% SDS, DTT, mercaptoethanol and other reducing agents,the salt concentrations above 50-100 mM.
Note
1. Enzymes should be stored in an ice box or on an ice bath when used, and should be stored at -20°C immediately after use.
2. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.
HB230406
Q：是无菌的产品吗？
A：是的。
[1] Lu Z, Liu H, Song N, et al. METTL14 aggravates podocyte injury and glomerulopathy progression through N6-methyladenosine-dependent downregulating of Sirt1. Cell Death Dis. 2021;12(10):881. Published 2021 Sep 27. doi:10.1038/s41419-021-04156-y(IF:8.469)
[2] Yu Y, Wang Y, Zhang W, et al. Biomimetic periosteum-bone substitute composed of preosteoblast-derived matrix and hydrogel for large segmental bone defect repair. Acta Biomater. 2020;113:317-327. doi:10.1016/j.actbio.2020.06.030(IF:7.242)