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Cat. No.: BRD-100 (for 100T)

Cat. No.: BRD-500 (for 500T)

Description

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

DNases and ribonucleases (RNases) are widely present in the environment and biological organisms. Due to their ability to degrade nucleic acids, the presence of nucleases can interfere with many experiments [1]. In processed biological products, trace amounts of these nucleases can have an impact on the subsequent applications of the products. Broad-spectrum nucleases such as Benzonase and Broadonase are commonly used non-specific endonucleases for the removal of nucleic acids in various biological products, including recombinant proteins and viral vaccines. Therefore, the residual levels of Benzonase, Broadonase, and other broad-spectrum nucleases in biological products are important indicators of product quality. The methods commonly described in the literature for detecting Benzonase, Broadonase, and similar nucleases are often time-consuming and have relatively low detection sensitivity.

This assay kit provides high sensitivity and can detect Benzonase or Broadonase at levels as low as approximately 0.002 U (approximately 0.003 ng). Based on the detection system calculation, the concentration of Benzonase or Broadonase in the sample is approximately 0.0002 U/μl or 0.3 pg/μl. This assay kit is also known as the Broadonase Nuclease Residue Detection Kit, Super Nuclease Residue Detection Kit, Super Nuclease Activity Fluorometric Detection Kit, Benzonase Activity Fluorometric Detection Kit, Fluorescence-based Super Nuclease Activity Detection Kit, Universal Nuclease Residue Detection Kit, or Broad-Spectrum Nuclease Residue Detection Kit.

Benzonase Residue Detection Kit utilizes the fluorescence resonance energy transfer (FRET) method for detection, as illustrated in Figure 1. The Benzonase substrate is a synthetic DNA oligonucleotide probe with a VIC fluorophore (donor) at one end and a BHQ1 quencher (acceptor) at the other end. These two fluorophores have overlapping absorption spectra. When the distance between the fluorophores is appropriate, the 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 attached to the ends of the Benzonase substrate. When the substrate is cleaved by Benzonase, the ends of the DNA substrate separate, causing the two fluorophores to dissociate. As a result, the fluorescence of VIC is no longer quenched by BHQ1, allowing the detection of VIC fluorescence. This fluorescence-based detection enables highly sensitive measurement of Benzonase nuclease activity. The maximum excitation wavelength of VIC is 535 nm, and the maximum emission wavelength is 556 nm. The kit includes a Benzonase standard, which can be used to set up a standard curve for calculating the residual amount of Benzonase in the samples.

Content

Cat. No.: BRD-100 (for 100T)

• 10X Reaction Buffer                2ml
• Standard (250U/μl)                   5μl
• 5X Benzonase Substrate    200μl
• Nuclease-free Water            20ml

Cat. No.: BRD-500 (for 500T)

• 10X Reaction Buffer               10ml
• Standard (250U/μl)                 20μl
• 5X Benzonase Substrate         1ml
• Nuclease-free Water            100ml

Features

• Benzonase Residue Detection Kit offers high sensitivity and requires a small sample volume. The kit includes Benzonase as a standard, facilitating the establishment of the detection system. The Benzonase substrate probe has been optimized to achieve high sensitivity, capable of detecting Benzonase as low as approximately 0.002 U (approximately 0.003 ng). Based on the detection system calculation, the concentration of Benzonase in the sample is approximately 0.0002 U/μl or 0.3 pg/μl. The detection sensitivity of this kit exceeds that of conventional products in the same category.
• Benzonase Residue Detection Kit offers a wide range of applications, providing flexibility and fast detection. This kit is not only suitable for detecting Benzonase but also other DNases, including nucleases that can cleave single-stranded and double-stranded DNA. Compared to ELISA methods, this kit offers a simpler, faster, and more accurate detection process. Experimental validation has confirmed that this kit can be used to detect the levels of super nucleases like Broadonase, DNase I, T4 DNA polymerase, T5 Exonuclease, Micrococcal nuclease, Mung Bean Nuclease, S1 Nuclease, and other enzymes. The kit utilizes a one-step detection method that is simple and fast, completing the entire process in approximately 10-20 minutes.

When used for 96-well plate assays, the small package of the kit allows for 100 tests, while the medium package enables 500 tests to be performed.

Precautions

• Due to the high sensitivity of this assay kit and the potential presence of nucleases in the environment, it is recommended to perform Benzonase residue detection in a clean environment such as a laminar flow hood or biosafety cabinet to prevent interference from nucleases present in the surroundings.
• The 10X Reaction Buffer, Nuclease-free Water, and 5X Benzonase Substrate should be fully thawed and equilibrated to room temperature before use, as failure to do so may affect the assay results. The Standard (250U/μl) should be kept on ice during use and all reagents should be stored according to the kit's instructions after use.
• Ensure that the sample pH is between 7-8 or adjust the pH of the reaction system after adding the sample, as deviations from this range may affect the signal values and stability of the assay results. For small 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 fully melted and mixed before use.
• This assay kit may not be suitable for certain nucleases, such as Klenow Fragment and Phi29 DNA polymerase. Generally, the provided 10X Reaction Buffer is compatible with most nucleases; however, there may be exceptions where specific nucleases require dilution and reaction in their respective buffers. Please dilute and react the samples with specific nucleases accordingly if necessary.
• To prevent DNase contamination of the reagents, it is advisable to use RNase and DNase Away to remove any DNases present in the environment before each experiment, if necessary.
• It is recommended to use 96-Well Black Opaque Plates for the assay. 
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, or in food or drug applications. It should not be stored in regular residential areas.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling the reagents.

Storage

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