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Deoxyribonuclease I (DNase I) GMP-grade (2 U/μL)
产品货号:10611ES76
产品规格:500 U
产品价格:¥ 645
类别:体外转录系列

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  • Product Information

     

    Product Name

    Catalog No.

    Size

    Price ()

    Deoxyribonuclease I (DNase I) GMP-grade (2 U/μL)

    10611ES76

    500 U

    645

    10611ES84

    2,000 U

    1965

    10611ES92

    10,000 U

    6455

     

    Product Description

     

    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 7.62 mM CaCl250%v/vGlycerol 

    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

    10611

    Deoxyribonuclease I (DNase I) GMP-grade (2 U/μL)

    250 μL

    1 mL

    5 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

    *10×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 2.5mM 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.

     

    HB220514