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Molecular Probes™ FISH Tag™ RNA Kits with Alexa Fluor™ Dyes
Description
Choose from two, single-color FISH Tag RNA kits with Alexa Fluor dye options (green or red). The FISH RNA multicolor kit contains four spectrally distinct dyes (green, orange, red, and deep-red) to allow simultaneous localization of multiple sequence-specific RNA and DNA targets such as mRNAs in cells, tissues, and whole embryo mounts. All kits include required reagents and come with a comprehensive, detailed protocol to help ensure your success.
Complete workflow solution for FISH
FISH Tag kits employ a two-step labeling technology. First, in vitro transcription is used to enzymatically incorporate an amine-modified nucleotide into the probe template. The modified nucleotide is UTP with an NH2 group attached through a linker to the C5 position of the base. Next, dye labeling of the purified amine-modified RNA is achieved by incubation with amine-reactive dyes (e.g., Alexa Fluor dyes). These active ester compounds react with the primary amines incorporated into the probe template, covalently conjugating the dye to the modified nucleotide base. The purified probe is then ready for hybridization to the specimen. Once the hybridization assay is complete, the specimen is viewed under a fluorescence microscope to visualize the hybridized fluorescent probe.
The dyes available in the FISH Tag RNA kits are compatible with standard filter sets found on most fluorescence microscopes. Alexa Fluor dyes are brighter and more photostable than traditional fluorescent labels, providing higher resolution and improved signal-to-noise ratios compared to conventional dyes. PureLink nucleic acid purification technology is used for fast and efficient purification of the labeled probe while SlowFade Gold Antifade Mountant preserves the fluorescent signal during imaging.
See the user manual for information regarding how to calculate base:dye ratio and nucleic acid concentration, how to construct plasmids using the FISH Tag RNA kits, and about optional RNA probe fragmentation for RNA probes greater than 500 bases.
Specifications
Specifications
| Content And Storage | Store in freezer (-5°C to -30°C) and protect from light. |
| Color | Red |
| Detection Method | Fluorescence |
| Format | Kit |
| Label or Dye | Alexa Fluor 594 |
| Final Product Type | Probes (Labeled RNA), Probes (Labeled DNA), Oligos (Labeled) |
| Sample Type | RNA, Tissue (FFPE), Cell Culture Cells, mRNA |
| Labeling Target | RNA (General), siRNA |
| Excitation/Emission | 590/615 nm |
| For Use With (Application) | FISH Tag, fluorescent microscopy, RNA In situ hybridization |
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Frequently Asked Questions (FAQs)
FISH probes are generally from 300 to 1500 bp in length. To adequately image a gene or other target, the FISH probes must cover a contiguous length of ˜10 kbp. Shorter strand lengths or low-abundance DNA or RNA targets may be detected with the use of fluorescent amplification methods, such as tyramide signal amplification (TSA). For more information using TSA to amplify FISH detection, see this manual (http://tools.thermofisher.com/content/sfs/manuals/mp20912.pdf).
RNA FISH probes are the best option for hybridization to mRNA, rRNA, or other RNA targets. DNA FISH probes can also be used to detect RNA targets, but they do not hybridize as well to mRNA and other RNA target molecules. DNA FISH probes are routinely used to detect gene loci on chromosomes. RNA FISH probes can also be used to detect genes on chromosomes, but DNA FISH probes are more robust for this application.
For Research Use Only. Not for use in diagnostic procedures.
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