Recombinant Deoxyribonuclease I  (DNase I,RNase-free) GMP-grade

Recombinant Deoxyribonuclease I (DNase I, RNase-free) GMP-grade (2 U/μL) is expressed from E. coli, widely used in the preparation of DNA-free RNA and removing the template DNA after in vitro transcription. Different from DNase I (Cat#10325), this GMP-grade DNase I is produced in accordance with GMP process requirements, and the product is provided in liquid form.

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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 7.6,2 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)

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

[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 the 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 a 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.