For general laboratory use. Others FastStart Essential DNA Probes Master GLU FastStart Essential DNA Probes Master 3.8.1.6.3.3 FastStart Essential DNA Probes Master 04038377028736 1 kit false FS Essential DNA Probes Master,10x5 ml 06924492001 10 x 500 reactions of 20 μL final volume each Reagents, kits FastStart Essential DNA Probes Master 04038377028088 1 kit false FastStart Essential DNA Probes Master 06402682001 500 reactions of 20 μL final volume each Reagents, kits en Sequence-specific detection of PCR products relies on sequence-specific oligonucleotide probes that are coupled to fluorophores. These probes hybridize to their complementary sequence in target PCR products. Hydrolysis probe chemistry uses the so-called FRET principle. Fluorescence Resonance Energy Transfer (FRET) is based on the transfer of energy from one fluorophore (the donor or reporter) to another adjacent fluorophore (the acceptor or quencher).Hydrolysis probe assays can technically be described as homogeneous 5'-nuclease assays, since a single 3' non-extendable probe, which is cleaved during PCR amplification, is used to detect the accumulation of a specific target DNA sequence. This single probe contains two labels, a fluorescent reporter and a quencher, in close proximity to each other. When the probe is intact, the quencher dye is close enough to the reporter dye to suppress the reporter fluorescent signal (fluorescent quenching takes place via FRET). During PCR, the 5'-nuclease activity of the polymerase cleaves the hydrolysis probe, separating the reporter and quencher. The reporter dye is no longer quenched and emits a fluorescent signal when excited.The LightCycler® PRO and the LightCycler® 96 Instrument are factory calibrated for commonly used reporter dyes as listed in the respective systems. For example, different labeled hydrolysis probes can be used separately or in combination, which permits up to four-color (LightCycler® 96 Instrument) or up to seven-color (LightCycler® PRO Instrument) detection. There is no need for color compensation runs.For multicolor hydrolysis probe assays, use dark quencher dyes, that is, dye molecules which efficiently quench the fluorescence of a FRET reporter dye without emitting fluorescence themselves. For example, use BHQ-2 (quenching range 550 to 650 nm) for all hydrolysis probe reporter dyes.Color compensation is automatically performed (all analysis data are color compensated).How this product worksFastStart Essential DNA Probes Master is a ready-to-use reaction mix specifically developed for the hydrolysis probe detection format in:LightCycler® 480 Multiwell Plates 96 or,8-Tube Strips without an adapter plate on the LightCycler® 96 Instrument or,LightCycler® 480 Multiwell Plates 96 or,LightCycler® 480 Multiwell Plates 384 or,8-Tube Strips with the LightCycler® 8-Tube Strip Adapter Plate on the LightCycler® PRO Instrument.The master contains FastStart Taq DNA Polymerase for hot start PCR, which significantly improves the specificity and sensitivity of PCR by minimizing the formation of nonspecific amplification products.FastStart Taq DNA Polymerase is a chemically modified form of thermostable recombinant Taq DNA Polymerase that shows no activity up to +75°C. The enzyme is active only at high temperatures, where primers no longer bind non-specifically. The enzyme is completely activated (by removal of blocking groups) in a single pre-incubation step (+95°C, 5 to 10 minutes) before cycling begins. Activation does not require the extra handling steps typical of other hot start techniques. en The FastStart Essential DNA Probes Master is a ready-to-use hot start reaction mix designed for detecting DNA targets with hydrolysis probes. It allows very sensitive detection and quantification of defined DNA sequences as well as endpoint genotyping analysis. The kit may also be used in other types of PCR on the LightCycler® PRO and LightCycler® 96 Systems.The kit can also help prevent carryover contamination during PCR (when used with LightCycler® Uracil-DNA Glycosylase) or to perform the second step of a two-step RT-PCR.In principle, the FastStart Essential DNA Probes Master can be used to amplify and detect any DNA or cDNA target. However, the detection protocol must be adapted to the reaction conditions of the LightCycler® PRO or LightCycler® 96 Instruments, and specific PCR primers and probes must be designed for each target. en FastStart Essential DNA Probes Master is a ready-to-use reaction mix containing FastStart Taq DNA Polymerase for hot start PCR, which significantly improves the specificity and sensitivity of PCR by minimizing the formation of nonspecific amplification products. The 2x master mix is optimized for a fixed MgCl2 concentration, which works with nearly all primer combinations.No adjustment in the MgCl2 concentration is needed to amplify different sequences; only template DNA, PCR primers, and hydrolysis probes must be added.