LightCycler® FastStart DNA Master HybProbe
Easy-to-use hot start reaction mix for PCR using the LightCycler® Carousel-Based System.
For general laboratory use.
Easy-to-use Hot Start Reaction Mix for PCR, using HybProbe probes with the LightCycler® Carousel-Based System.
LightCycler® FastStart DNA Master HybProbe provide convenience, high performance, and reproducibility in quantitative PCR.
- LightCycler® FastStart Enzyme
- LightCycler® FastStart Reaction Mix HybProbe
- MgCl2 Stock Solution, 25 mM
- Water, PCR Grade
LightCycler® FastStart DNA Master HybProbe is an easy-to-use hot start reaction mix for sensitive PCR applications in LightCycler® capillaries, using HybProbe probes as detection format. It is an ideal master mix for performing quantitative PCR, as well as SNP and mutation detection and can also be used in two-step RT-PCR.
Note: LightCycler® FastStart DNA MasterPLUS HybProbe can be used in conjunction with Uracil DNA Glycosylase, heat-labile for carryover prevention during PCR.
The unique LightCycler® HybProbe format is based on the principle of fluorescence resonance energy transfer (FRET). Two sequencespecific oligonucleotide probes, labeled with different dyes (donor and acceptor), are added to the reaction mix in addition to PCR primers. HybProbe probes hybridize to the target sequences on the amplified DNA fragment during the annealing phase in a head-totail arrangement, thereby bringing the two dyes into close proximity. The donor dye (fluorescein) is excited by the blue light LED source and the energy emitted by the donor dye excites the acceptor dye attached to the second HybProbe probe, which then emits fluorescent light at a different wavelength. The amount of fluorescence is directly proportional to the amount of target DNA generated during the PCR process.
The hot start feature of the master mixes is provided by FastStart Taq DNA Polymerase, a chemically modified Taq DNA polymerase, which is inactive at room temperature and becomes activated at high temperatures (i.e., a preincubation step at +95°C for 5 – 10 minutes). If necessary, the MgCl2 concentration can be optimized for specific applications.
Note: All LightCycler® Master Mixes have been specifically developed to function in LightCycler® Capillaries and provide optimal performance and sensitivity.
Function test: The LightCycler® FastStart DNA Master HybProbe is function tested using the LightCycler® Control Kit DNA, according to the kit protocols.