DNase Alert Kit
Deoxyribonuclease (DNase) is an enzyme that is ubiquitous in the environment and in many biological materials. Because DNase can degrade DNA, its presence poses a threat to many molecular biology experiments. It is necessary to develop fast and sensitive DNA detection methods. The DNase Alert Assay is a fast and convenient test that can detect DNase contamination in 10 minutes or less. The DNase Alert Kit System contains a unique substrate that is labeled with both fluorescent reporter and quenching groups. Due to the close proximity of the fluorophores and quenched groups, the fluorescence emitted by the unlyzed substrate is very weak. When DNase is present, the chemical bonds between the fluorophore and quenching group are broken, resulting in a bright green signal when excited at the appropriate wavelength. The intensity of fluorescence is proportional to the degree of substrate cleavage, which is proportional to the degree of DNase contamination.This kit can be used to detect DNase I and other DNA nucleases that cleave double-stranded DNA, as well as enzymes that cleave single-stranded DNA, such as exonuclease III, mung bean nuclease, micrococcal nuclease, Bal31 nuclease, S1 nuclease, T7 endonuclease, etc. Most reaction buffers and solutions for DNA can be detected with this kit, but some systems are not compatible, such as (1) gel loading buffer and other dark solutions, as dark solutions may interfere with the excitation of fluorescence or prevent its emission. (2) Solutions that inhibit DNase activity. Known common DNase inhibition solutions include high ionic strength solutions (e.g., 5 M NaCl, 20 X SSC, 3 M sodium acetate, etc.), solutions with pH less than 4 or greater than 9, dispersants, detergents, chelating agents, or any solution that denatures proteins (e.g., SDS, guanidine thiocyanate, urea, EDTA, etc.); (3) Solutions that cause chemical instability of DNase Alert substrates. DNase Alert substrates are unstable in solutions with pH >9 and in corrosive solutions (strong acids, bases, bleach).
| Components | K1901-100 T | K1901-500 T |
| DNaseAlert Substrate (2 nmol/tube) | 1 tube | 5 X 1 tube |
| 10X NucleaseAlert Buffer | 2.5 mL | 11 mL |
| DNase I (2 U/μL) | 10 μL | 50 μL |
| TE Buffer (pH 7.0) | 1.5 mL | 6 mL |
| Nuclease-free Water | 10 mL | 50 mL |
Store Nuclease-free Water at room temperature, DNaseAlert Substrate the components at -20°C away from light, and the others at -20°C for 12 months. | ||