For life science research only. Not for use in diagnostic procedures. Others SimpleProbe 519 Labeling Reagent LSR SimpleProbe 519 Labeling Reagent 15 70051125019 This product is sold for use by purchaser only. The sale or distribution of this product or any product derived from the use of this product to any third party is expressly excluded. Especially excluded is the commercial use or sale of oligonucleotides or other products labeled with this product. 3.5.8.2.1.2 04687132001 Simple Probe 519 Labeling Reagent SimpleProbe 519 Labeling Reagent 04038377021805 Reagents, kits 100 μmol For preparation of 10 internally labeled oligonucleotides (0.2 μmol scale) 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. en 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. en 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.   en