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

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

Contents

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:

【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