DNaseAlertTM Substrate and Nuclease Detection System Nucleases are widely present in the laboratory environment and can interfere with many experiments. IDT has developed reagents that allow for rapid, sensitive detection of RNases and DNases. These reagents are fluorescence-quenched oligonucleotide probes that emit a fluorescent signal only after nuclease degradation. The assay can be read visually or measured and quantified using fluorometry. Assays can be used qualitatively to test lab reagents, equipment and supplies for nuclease contamination. Assays can also be used quantitatively to study enzyme kinetics. The DNaseAlertTM Substrate is a synthetic DNA oligonucleotide that has a HEXTM reporter dye (hexachlorofluorescein) on one end and a dark quencher on the other end. The DNA sequence has been carefully optimized to react with a wide variety of nucleases; it contains domains that will react with single-stranded endonucleases, certain single-stranded exonucleases, and double-stranded nucleases. Intact the substrate has little or no fluorescence. When cleaved by a DNase, the substrate fluoresces pink (536 nm or UV excitation, 556 nm emission) and can be detected visually or using a fluorometer. DNaseAlertTM is available in single-use tubes which are ideal for rapid hand-held visual assays at the research bench, or as bulk substrate which can be used in either visual assays or read in a fluorometer. DNaseAlertTM is also available as a kit that provides 25 single-use tubes of the DNaseAlertTM Substrate. The kit also includes a buffer optimized for detection of a wide variety of DNases, nuclease-free water, DNase 1 Enzyme (for use as a positive control), and 50 ml of Nuclease Decontamination Solution (to clean lab surfaces found to have nuclease contamination). |