All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: DIRD-100 (for 100T)

Cat. No.: DIRD-500 (for 500T)

Description

DNase I Residue Detection Kit or Fluorometric DNase I Assay Kit is a reagent kit used for the rapid and highly sensitive detection of DNase I and other deoxyribonucleases (DNases) residue in samples using fluorescence-based methods.

Similar to ribonucleases (RNases), DNases and RNases are widely present in experimental environments and biological organisms. Due to their ability to degrade nucleic acids, the presence of these enzymes can interfere with many experiments. The trace amounts of these nucleases remaining in processed biological products can have an impact on the subsequent application of the products. Common methods for detecting DNase I in the literature are often time-consuming and have relatively low sensitivity.

This kit offers high detection sensitivity and can detect DNase I as low as approximately 6×10-4 U. The kit is also referred to as the DNase I Residue Detection Kit, DNase Residue Detection Kit, DNase I Activity Fluorometric Assay Kit, DNAse I Activity Fluorometric Assay Kit, or Fluorescence-based DNase I Activity Assay Kit.

DNase I, also known as deoxyribonuclease I, is the most common DNase. It is an endonuclease that can digest single-stranded or double-stranded DNA, producing single deoxyribonucleotides or short oligodeoxyribonucleotides. The hydrolysis products of DNase I on single-stranded or double-stranded DNA have a phosphate group at the 5' end and a hydroxyl group at the 3' end. The activity of DNase I depends on calcium ions and can be activated by magnesium ions or divalent manganese ions. In the presence of magnesium ions, DNase I can randomly cleave double-stranded DNA at any position. In the presence of divalent manganese ions, DNase I can cleave the DNA double helix at the same site, resulting in blunt ends or sticky ends with 1-2 protruding nucleotides.

The DNase I Residue Detection Kit utilizes the principle of fluorescence resonance energy transfer (FRET). The DNase substrate is a synthetic DNA oligonucleotide probe that has a VIC fluorophore (donor) at one end and a BHQ1 quencher (acceptor) at the other end. These two fluorophores have partially overlapping absorption spectra. When the distance between these two fluorophores is appropriate, fluorescence energy is transferred from the donor to the acceptor, resulting in a decrease in the fluorescence intensity of the donor fluorophore. VIC and BHQ1 are connected to the ends of the DNase I substrate. After the substrate is cleaved by DNase I, the ends of the DNA substrate separate, causing the two fluorophores to dissociate. The fluorescence of VIC can then be detected, indicating the activity of DNase enzyme with high sensitivity. The maximum excitation wavelength of VIC is 535nm, and the maximum emission wavelength is 556nm. The kit provides a DNase I standard, which can be used to calculate the residual amount of DNase I in the sample by setting a standard curve. The kit can also detect other non-DNA sequence-specific DNA endonucleases.

Content

Cat. No.: DIRD-100 (for 100T)

• 10X Reaction Buffer 2ml 
• DNase I (1U/μl) 20μl 
• 5X DNase I Substrate 200μl 
• Nuclease-free Water 20ml

Cat. No.: DIRD-500 (for 500T)

• 10X Reaction Buffer 10ml 
• DNase I (1U/μl) 100μl 
• 5X DNase I Substrate 1ml 
• Nuclease-free Water 100ml

Features

• This assay kit offers high detection sensitivity and requires a small sample volume. The kit provides DNase I as a positive control, facilitating the establishment of the detection system. Furthermore, the DNase I substrate probe has been optimized for high sensitivity, allowing the detection of DNase I as low as approximately 6×10-4 U. The detection sensitivity of this kit exceeds that of conventional similar products.
• This assay kit has a wide range of applications, flexible usage, and fast detection speed. It can not only be used to detect DNase I but also other DNases, including nucleases that can cleave single-stranded and double-stranded DNA. Compared to ELISA methods, this kit is simpler, faster, and more accurate. Experimental validation has shown that this assay kit can be used to detect the relative activity of enzymes such as Benzonase-like super nucleases, T4 DNA polymerase, T5 exonuclease, Micrococcal nuclease, Mung Bean nuclease, and S1 nuclease. The assay kit utilizes a one-step detection method, which is simple and rapid, requiring only approximately 20 minutes to complete the entire process.

When used for 96-well plate detection, the small package of this assay kit can perform 100 tests, and the medium package can perform 500 tests.

Precautions

• Due to the interference of DNase present in the environment, it is recommended to perform DNase I residue detection in a relatively clean environment such as a laminar flow hood or a biosafety cabinet to prevent the influence of environmental DNase on the test samples.
• The 10X Reaction Buffer, Nuclease-free Water, and 5X DNase I Substrate should be completely thawed and equilibrated to room temperature before use, as failure to do so may affect the test results. DNase I (1U/μl) should be kept on ice during use and all reagents should be stored according to the kit's requirements immediately after use.
• Please ensure that the pH value of the sample is between 7 and 8 or that the pH value of the reaction system falls within this range after adding the sample, as it may affect the signal values and stability of the test results. For smaller volume reagents, it is recommended to briefly centrifuge them to ensure the liquid settles at the bottom of the tube before use. Thawed reagents must be completely melted and mixed well before use.
• This kit may not be suitable for certain nucleases, such as Klenow Fragment and Phi29 DNA polymerase. In general, the provided 10X Reaction Buffer in this kit is compatible with most nucleases, but there may be exceptions for specific enzymes. If necessary, dilution and reaction of the samples with specific nuclease buffers are recommended.
• During the use of this kit, precautions should be taken to prevent contamination of the reagents by DNase. If necessary, RNase and DNase Away can be used to remove DNase present in the environment.
• It is recommended to use 96-Well Black Opaque Plates for detection.
• This product is intended for scientific research purposes by professionals only and is not to be used for clinical diagnosis or treatment, food or drug purposes, or stored in ordinary residences.
• For your safety and health, please wear lab coat and disposable gloves while performing the experiments.

Storage

The minimum shelf life is 1 year at -20°C. The 5X DNase I Substrate should be stored protected from light.