LightCycler® RNA Amplification Kit HybProbe
Kit for one-step RT-PCR using the LightCycler® Carousel-Based System.
For general laboratory use.
The LightCycler® RNA Amplification Kit HybProbe provides reagents, including enzyme mix, reaction mix, MgCl2, and PCR-grade water for amplification and detection of RNA using the LightCycler® Carousel-Based System Instruments.
The LightCycler® RNA Amplification Kit HybProbe product provides convenience, high performance, reproducibility, and minimized contamination risk. Addition of MgCl2 (provided in the kit) may be necessary to optimize performance.
- LightCycler® RT-PCR Enzyme Mix
- LightCycler® RT-PCR Reaction Mix HybProbe
- MgCl2 Stock Solution, 25 mM
- Water, PCR-grade
The LightCycler® RNA Amplification Kit HybProbe is specifically adapted for one-step RT-PCR in LightCycler® Capillaries using the LightCycler® Carousel-Based System Instruments and HybProbe probes as detection format.
HybProbe probes consist of two different oligonucleotides that hybridize to an internal sequence of the amplified fragment during the annealing phase of PCR. One probe is labeled at the 5' end with a LightCycler® Red fluorophore (LC Red 640 or LC Red 705), and to avoid extension, modified at the 3' end by phosphorylation. The other probe is labeled at the 3' end with fluorescein. Only after hybridization to the template, the two probes come in close proximity, resulting in fluorescence resonance energy transfer (FRET) between the two fluorophores. During FRET, fluorescein, the donor fluorophore, is excited by the light source of the LightCycler® Instruments. Part of the excitation energy is transferred to LightCycler® Red, the acceptor fluorophore. The emitted fluorescence of the LightCycler® Red fluorophore is measured.
Function test: The LightCycler® RNA Amplification Kit HybProbe is function tested using the LightCycler® Control Kit RNA, according to the kit protocols.