膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | FRET |
| Analyte | DNase I |
| Format | 96T/480T |
| Regulatory Status | RUO |
| Sensitivity | 3.9×10⁻⁵ U |
| Standard Curve Range | 0.0025 U-3.9×10⁻⁵ U |
| Assay Time | 30 min |
| Suitable Sample Type | For the quantitative determination of DNase I in the environment, some biological materials, common molecular biological reagents such as reaction buffers. |
| Sample volume | 10 μL & 80 μL |
组分（Materials Provided）
| Items | Size （96tests） | Size（480tests） |
| DNase Substrate | 2 nmol | 10 nmol |
| 10X Reaction Buffer for DNase | 10 mL | 10 mL |
| DNase I (1U/μL) | 20 μL | 50 μL |
| TE Buffer (pH 8.0) | 1.5 mL | 6 mL |
| Nuclease-free Water | 10 mL | 50 mL |
背景（Background）
Deoxyribonucleases (DNases) are enzymes which are able to hydrolyze phosphodiester bonds of DNA molecules. They can be divided into two families, which differ in biochemical and biological properties—DNase I and DNase II families. The presence of DNases would affect many experimental results like PCR, so it is necessary to evaluate the presence of DNases.
DNases are ubiquitous in both the environment and many biological materials. Many molecular biology experiments rely on the use of plastics, chemicals, and solutions that are free from detectable DNase activity, once these materials are contaminated with DNases, the experiments will be affected because of the DNases can degrade DNA. Since even only minute amounts of DNase contamination would ruin the experiment, it is necessary to evaluate the presence of DNase with a reasonable method.
Published methods for detecting DNase such as Nucleic acid hydrolyzed gel electrophoresis and ultraviolet spectrophotometer are typically time consuming, not quantitative, and relatively insensitive. The other methods such as HPLC and electrochemical methods are limited to instrumentation.
In contrast, the DNase Activity Assay Kit (Fluorescence) can detect DNase contamination within 30 minutes, and the kit is high sensitivity, easy to use. Moreover, the DNase and RNase Activity Assay Kit (Fluorescence) have been designed to work together seamlessly for simultaneous quantitative detection of DNases and RNases in a single sample.
应用说明（Application）
The DNase Activity Assay Kit (Fluorescence) is a convenient and sensitive assay tool to test the presence of DNase in buffers, reagents, and other components.
It is for research use only.
重构方法（Reconstitution）
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
The unopened kit should be stored at -20°C upon receiving.
Find the expiration date on the outside packaging and do not use reagents past their expiration date.
The opened kit should be stored per components table. The shelf life is 3 months from the date of opening.
原理（Assay Principles）
The DNase Activity Assay Kit (Fluorescence) is based on a fluorophore-labeled DNase substrate. When the sample does not contain DNase activity, the substrate is stable and does not produce a fluorescent signal; when the sample contains DNase activity, the substrate is degraded, resulting in a gradual enhanced fluorescence signal, the rate of increase in fluorescence signal is positively correlated with the number and activity of enzymes. Use a fluorescence microplate reader to measure at the wavelength of ex/em = 535/565 nm to determine whether the sample is contaminated by DNase.
活性（Bioactivity）-Fluorescence Please refer to Ds document for the assay protocol.
Add 90 μL of the working DNase Substrate solution to each 96-well plate, and add 10 μL of DNase I standards (0.00000390625-0.00025 U/μL*10μL/well = 0.0000390625-0.0025U/well), incubate the plate in the fluorometer (BMG CLARIOstar) collecting real-time data at 1 minute intervals for 30 minutes at 37°C using the settings described in this section. The DNase Activity Assay can be evaluated in rigorous kinetic terms using real-time data.
This assay kit employs a standard detection of DNase I. Add 90 μL working DNase Substrate solution to each 96-well plate, and add 10 μL DNase I standards ((0.00000390625-0.00025 U/μL*10μL/well = 0.0000390625-0.0025U/well), incubate for 30 minutes at 37°C. Then measure the fluorescence using the settings described in this section in a fluorometer (BMG CLARIOstar). Take RFU of standards as the ordinate and DNase concentration as the abscissa. Four parameters logistic are used to draw the standard curve (QC tested).
验证（Validation）
批内差异（Intra-Assay Statistics）
Eight replicates of each of seven samples containing different concentrations of DNase were tested in one assay, Intra-Assay Precision CV＜10%.
批间差异（Inter-Assay Statistics）
Eight replicates of each of seven samples containing different concentrations of DNase were tested in eight independent assays, Inter-Assay Precision CV<15%.
稳定性 (Stability)
The probe is freeze-thawed 3 times, the kit performance meets the requirements, the sensitivity is not reduced, and the CV< 10%.
回收率（Recovery）
Three different concentrations of DNase was spiked into four different kinds of matrixes. The range of the recovery rate is 80%~120%.
干扰效应(Interference effect)
DNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A002) and RNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A001) were used to detect nuclease residues in the same sample. No significant cross-reactivity or interference was observed.
