Description
The FluoroStain™ DNA Fluorescent Staining Dye is designed to be a safer replacement for conventional Ethidium bromide (EtBr) which poses a significant health and safety hazard for its users. The FluoroStain™ DNA Fluorescent Staining Dye offers at least 10 times sensitivity in DNA detection levels, and is capable of detecting double stranded DNA (dsDNA) fragments up to 0.04 ng in electrophoresis analysis. The FluoroStain™ DNA Fluorescent Staining Dye shows a high specificity to the dsDNA, with negligible background signal, making the destaining process entirely optional. FluoroStain™ DNA Fluorescent Staining Dye is compatible with both the conventional ultra violet gel-illuminating systems as well as the less harmful long wave length blue light illumination systems. The emission when bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
Features:
Storage
Protected from light
4°C for 12 months
-20°C for 24 months
The FluoroStain™ DNA Fluorescent Staining Dye is designed to be a safer replacement for conventional Ethidium bromide (EtBr) which poses a significant health and safety hazard for its users. The FluoroStain™ DNA Fluorescent Staining Dye offers at least 10 times sensitivity in DNA detection levels, and is capable of detecting double stranded DNA (dsDNA) fragments up to 0.04 ng in electrophoresis analysis. The FluoroStain™ DNA Fluorescent Staining Dye shows a high specificity to the dsDNA, with negligible background signal, making the destaining process entirely optional. FluoroStain™ DNA Fluorescent Staining Dye is compatible with both the conventional ultra violet gel-illuminating systems as well as the less harmful long wave length blue light illumination systems. The emission when bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
Norgen’s Urine Total RNA Purification Maxi Kit Dx (Slurry Format) provides a fast, reliable and simple procedure for isolating total RNA from urine for subsequent in vitro diagnostic use. Purification is based on the use of Norgen’s proprietary resin as the separation matrix, and the kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to small RNAs.
This kit is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of RNA followed by signal detection or amplification. Any diagnostic results generated using the RNA isolated with Urine Total RNA Purification Maxi Kit Dx (Slurry Format) in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings.
To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used.
Norgen’s Urine Total RNA Purification Maxi Kit Dx (Slurry Format) is intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques including RNA isolation.
Norgen’s Urine Total RNA Purification Maxi Kit Dx (Slurry Format) does not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.
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| Kit Specifications | |
| Volume of Urine Processed | 5 – 10 mL |
| Size of RNA Purified | All sizes, including < 200 nt |
| Time to Complete 10 Purifications | < 30 minutes |
| Average Yield | Variable depending on specimen |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable until the expiration date specified on the label.
| Component | Cat. Dx29600 (50 preps) |
|---|---|
| RNA Lysis Solution | 3 x 90 mL |
| RNA Wash Solution | 24 mL |
| RNA Elution Solution | 6 mL |
| Mini Filter Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Endonucleases DNA-specific, dsDNase
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
In figure 1, a PCR master mix was treated with different amounts of dsDNase before performing a qPCR to measure the contaminating bacterial DNA in the master mix. ArcticZymes dsDNase effectively removed contaminating DNA below known levels of the assay detection limits.
The dsDNase from Arctic shrimp (Pandalus borealis) is recombinantly produced in Pichia pastoris. It cleaves phosphodiester linkages in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.
The specific activity is estimated to be 30 times higher than that of bovine DNase I. In the presence of magnesium as only divalent cation and using oligos as a substrate, the activity towards dsDNA is 5000-fold higher than towards ssDNA.
The unique double strand-specificity allows specific degradation of dsDNA while leaving shorter ssDNA as primers and probes essentially intact. Easy inactivation by moderate heat (65°C) allows addition of DNA intended for analysis directly after removal of contaminating DNA.
Figure 1. The dsDNase effectively removes contaminated DNA
The dsDNase effectively removes contaminated DNA:
A PCR master mix was preincubated with various concentrations of dsDNase. After treatment, no DNA was amplified in non-template controls.
Nucleic acid specificity has been tested towards double- and single-stranded DNA and RNA oligonucleotides. The specificity of dsDNase towards the substrate has been measured using 15-mer oligonucleotides with FAM at 5′ and DarkQuencher® 3′ (Eurogentec). The fluorescence is proportional to enzyme activity. Assay conditions: 25 mM Tris pH 7.5, 5 mM MgCl2, and 2 μM oligonucleotide.
Substrate Relative Activity
dsDNA 100%
ssDNA <0.03%
dsRNA <0.01%
ssRNA <0.01%
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.