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For general laboratory use. Others LightCycler Control Kit DNA GLU LightCycler® Control Kit DNA 3.5.5.1.1.4 12158833001 LC Control Kit DNA LightCycler Control Kit DNA 04038377016696 Reagents, kits 1 kit 50 reactions with 20 μl final volume each The LightCycler® Control Kit DNA allows performance of a control reaction for PCR using the LightCycler® Carousel-Based System. A 110 bp fragment of the β-globin gene is amplified with specific primers from human genomic DNA. Amplification and online monitoring of the control gene is achieved by a combined procedure on the LightCycler® System. The results are interpreted directly after completing the PCR. The amplicon is detected by fluorescence using either the dsDNA intercalating dye SYBR Green I or a β-globin-specific pair of HybProbe probes provided in the kit.How this Product WorksThe LightCycler® Control Kit DNA allows control reaction for PCR to be performed using the LightCycler® System and the respective LightCycler® Kit (LightCycler® DNA Master, LightCycler® DNA FastStart Master, or LightCycler® DNA FastStart MasterPLUS).For detection using SYBR Green I, a 110 bp fragment of the human β-globin gene is amplified with specific primers in the presence of this DNA double-strand specific dye. During online monitoring, the fluorescence increases as a result of the increasing amount of PCR product. Identification of the correct PCR product is done by Melting Curve Analysis.For detection using HybProbe probes, a 110 bp fragment of the human β-globin gene is amplified with specific primers from human genomic DNA. Amplification of target DNA is monitored using HybProbe pairs that hybridize to an internal sequence of the amplified fragment. HybProbe probes are two different short oligonucleotides that bind to an internal sequence of the amplified fragment during the annealing phase of the amplification cycle. One probe is labeled at the 5´-end with a LightCycler® Red fluorophore LightCycler® Red 640 and Cy5.5; it is also 3´-phosphorylated, so it cannot be extended. The other probe is labeled at the 3´-end with fluorescein. When hybridized to the template DNA, the two probes are close enough to allow fluorescence resonance energy transfer FRET between the two fluorophores.During FRET, fluorescein the donor fluorophore is excited by the light source of the LightCycler® Instrument. Fluorescein transfers part of this excitation energy to the LightCycler® Red dye the acceptor fluorophore. Then, the LightCycler® Red dye emits fluorescence, which is measured by the LightCycler® Instrument. HybProbe probes that contain different LightCycler® Red labels can be used separately for single color detection experiments or combined for dual color detection experiments. Color compensation is not necessary for single color detection experiments. However, if you are using HybProbe probes to perform dual color experiments in a single capillary, you must also use a color compensation file. Color compensation may be applied either during or after a run on the LightCycler® Instrument. en The LightCycler® Control Kit DNA includes human genomic DNA as the control template, control primers, HybProbe probes and PCR-grade water for performing PCR using the LightCycler® Carousel-Based System. It is designed to confirm PCR performance using LightCycler® Carousel-Based Instruments. en The LightCycler® Control Kit DNA is designed for use in research studies and includes human genomic DNA as the control template, control primers, HybProbe mixes, and Water, PCR Grade, for performing PCR using the LightCycler® Instruments. It is designed as a control to confirm PCR system performance using the LightCycler® System or as a quality check of the human DNA used in a specific assay, when combined with the function-tested primers and HybProbe mixes provided with the kit.Alternatively, the kit can be used as a quality check of specific primers and HybProbe probes by using the human genomic DNA provided with the kit. Furthermore, the kit can be used as a functional check of dual-color experiments and color compensation when simultaneously using the provided LightCycler® Red 640- and Cy5.5-labeled HybProbe pairs. en