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nm", "Country": "XG", "Code": "Emission", "Name": "Emission" }, { "Language": "en", "Value": "Store the product at −15 to −25°C.", "Country": "XG", "Code": "Storage Conditions (Product)", "Name": "Storage Conditions (Product)" }, { "Language": "en", "Value": "Excitation maximum at 494 nm", "Country": "XG", "Code": "Excitation Maximum", "Name": "Excitation Maximum" }, { "Language": "en", "Value": "SimpleProbe chemistry uses a single, fluorescently labeled probe to hybridize to a target sequence containing the SNP of interest. Immediately following PCR, amplified product is denatured and rapidly cooled, allowing the SimpleProbe probe to hybridize to its target-specific sequence. Once hybridized, the SimpleProbe probe emits a greater fluorescent signal than it does when it is not hybridized to its target.
At low temperatures, the fluorescent signal from the hybridized probes is high due to the large number of probes hybridized to their target sequence. As the temperature of the reaction is slowly increased, the SimpleProbe probe melts at the Tm of the probe, causing the fluorescent signal to decrease. Changes in fluorescent signal depend solely on the hybridization status of the probe. Mutations, such as SNP, weaken the binding of the SimpleProbe probe, causing the probe to melt off at a lower Tm compared to the wild-type sequence.
SimpleProbe probes are excellent tools for SNP genotyping and mutation detection because they readily identify wild-type, mutant, and heterozygous samples using a single short probe.

Design of SimpleProbe probes
Use the LightCycler® Probe Design Software, free download at www.lifescience.roche.com.", "Country": "XG", "Code": "Background Information - Help Corner", "Name": "Background Information - Help Corner" }, { "Language": "en", "Value": "
Vial / Bottle Label Function / Description Content
1 SimpleProbe 519 Labeling Reagent
  • Contains SimpleProbe 519 Labeling Reagent.
  • The reagent is supplied as a yellow solid filled in glass bottles adaptable to oligonucleotide synthesizers.
1 bottle, 100 μmol
", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "
For Preparation of the SimpleProbe 519 Labeling Reagent Solution
  • Syringe/needle
  • Anhydrous acetonitrile
For Oligonucleotide Labeling and Phosphorylation
  • DNA synthesizer
  • Vacuum centrifuge
  • Standard reagents for oligonucleotide synthesis (tetrazole, etc.)
  • 30 – 33% ammonia solution
  • 3'-Phosphate-CPG-support
  • N6-Bz-dA-Phosphoramidite
  • N2-iBu-dG-Phosphoramidite
  • N4-Bz-dC-Phosphoramidite
  • T-Phosphoramidite
For Oligonucleotide Purification by HPLC
  • HPLC
  • Strong anion exchange material (for example, Mono Q HR, or Source 15 Q). This separation medium is recommended to obtain optimal purification results.
  • Photometer
  • Sodium hydroxide (10 mM; pH ∼12)
  • Sodium chloride (1 M)
For Quality Control of HPLC-Purified Oligonucleotide
  • Photometer
  • 1 M sodium borate buffer (pH 8.5)
", "Country": "XG", "Code": "Additional Equipment and Reagent Required", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "Oligonucleotides react with the SimpleProbe 519 Labeling Reagent during solid phase phosphoramidite synthesis. This reagent can be used in the same manner as standard phosphoramidite (with 2 minutes coupling time) since it contains a dimethoxytrityl group. The reagent is stable during oligonucleotide synthesis. HPLC purification is simple and results in a well-defined product.", "Country": "XG", "Code": "Product Description", "Name": "Product Description" }, { "Language": "en", "Value": "SimpleProbe 519 Labeling Reagent is used for internal labeling of a mutation detection probe, which could be used in the SimpleProbe detection format with the respective LightCycler® System.", "Country": "XG", "Code": "Applications", "Name": "Applications" }, { "Language": "en", "Value": "SimpleProbe chemistry uses a single, fluorescently labeled probe that is designed to hybridize to a target sequence containing the SNP of interest. Immediately following PCR, amplified product is denatured and rapidly cooled, allowing the SimpleProbe probe to hybridize to its target-specific sequence. Once hybridized, the SimpleProbe probe emits a greater fluorescent signal than it does when it is not hybridized to its target. At low temperatures, the fluorescent signal from the hybridized probes is high due to the large number of probes hybridized to their target sequence. As the temperature of the reaction is slowly increased, the SimpleProbe melts at the Tm of the probe, causing the fluorescent signal to decrease. Thus, changes in fluorescent signal depend solely on the hybridization status of the probe. Mutations, such as SNP, weaken the binding of the SimpleProbe probe, causing the probe to melt off at a lower Tm compared to the wild type sequence. Therefore, SimpleProbe probes are an excellent tool for SNP genotyping and mutation detection because they readily identify wild type, mutant, and heterozygous samples with only a single short probe.
Please find the LightCycler® Probe Design Software 2.0 on our website for designing optimized PCR primers and probes for a given DNA sequence.

Spectral Characteristics
An oligonucleotide internally labeled with SimpleProbe 519 Labeling Reagent shows an excitation maximum at 494 nm and an emission maximum at 519 nm (both measured in 50 mM Tris, pH 8.3).
The 3' end of the probe has to be phosphorylated to avoid elongation during PCR.
Recommended length of oligonucleotides is 20 - 30 nt.

 
", "Country": "XG", "Code": "Principle", "Name": "Principle" } ] } } ] }

SimpleProbe 519 Labeling Reagent

For the synthesis of labeled oligonucleotides

SimpleProbe 519 Labeling Reagent

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