For general laboratory use. Others LightCycler FastStart DNA MasterPLUS HybProbe GLU LightCycler® FastStart DNA MasterPLUS HybProbe 3.5.5.1.3.5 LightCycler FastStart DNA MasterPLUS HybProbe 04038377019642 1 kit false LC FS DNA Master PLUS Hy.Pb, 480 react. 03515567001 480 reactions of 20 μl final volume each Reagents, kits LightCycler FastStart DNA MasterPLUS HybProbe 04038377020297 1 kit false LC FS DNA MasterPLUS HP, 100 ul 03752178001 1,920 reactions of 20 μl or 384 reactions of 100 μl final volume each Reagents, kits LightCycler FastStart DNA MasterPLUS HybProbe 04038377019635 1 kit false LC FS DNA Master PLUS HY-Pb, 96 react. 03515575001 96 reactions of 20 μl final volume each Reagents, kits en 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 fluorophore dye (LightCycler® Red 610, LightCycler® Red 640, Cy5, or Cy5.5) and to avoid extension, modified at the 3' end by phosphorylation. 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 dye (the acceptor fluorophore). Then, the dye emits fluorescence, which is measured by the LightCycler® Instrument. HybProbe probes that contain different dye 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.LightCycler® Red 610 and Cy5 can be used on a LightCycler® 2.0 Instrument only.How this Product WorksLightCycler® FastStart DNA MasterPLUS HybProbe is a ready-to-use reaction mix designed specifically for the HybProbe probes detection format using the LightCycler® Carousel-Based System. It is used to perform hot start PCR in glass capillaries. Hot start PCR has been shown to significantly improve the specificity and sensitivity of PCR (Chou, Q., et al., 1992; Kellogg, D.E., et al., 1994; Birch, D.E., et al., 1996), by minimizing the formation of nonspecific amplification products at the beginning of the reaction.FastStart Taq DNA Polymerase is a modified form of thermostable recombinant Taq DNA polymerase that is inactive at room temperature and below. The enzyme is active only at high temperatures, where primers no longer bind nonspecifically. The enzyme is activated (by removal of blocking groups) in a single pre-incubation step (95°C, 10 minutes) before cycling begins. Activation does not require the extra handling steps typical of other hot start techniques.LightCycler® FastStart DNA MasterPLUS HybProbe provides convenience, excellent performance, reproducibility, and minimal contamination risk. All you have to supply is template DNA, PCR primers, and HybProbe probes.The reaction mix in this kit is optimized for a single MgCl2 concentration, which works with nearly all primer combinations. You do not need to adjust the MgCl2 concentration to amplify different sequences.In combination with the LightCycler® Carousel-Based System and suitable PCR primers, this kit allows very sensitive detection and quantification of defined DNA sequences. Various sources of DNA (cDNA, genomic DNA, plasmid DNA, etc.) can be used. The kit is also suitable for SNP and mutation detection using melting curve analysis. Furthermore, the kit can be used to perform two-step RT-PCR in combination with a reverse transcription kit for cDNA synthesis. The amplicon size should not exceed 1 kb in length. For optimal results, select a product length less than 700 bp.LightCycler® FastStart DNA MasterPLUS HybProbe offers convenience and ease of use because adaption of MgCl2 in the reaction mixture is not necessary, thus avoiding time-consuming optimization steps. The new buffer formulation results in increased PCR robustness. en LightCycler® FastStart DNA MasterPLUS HybProbe is designed for PCR applications using the LightCycler® Carousel-Based System. The kit is ideally suited for hot start PCR applications.The kit can also be used in conjunction with heat-labile Uracil-DNA Glycosylase to prevent carryover contamination during PCR.