LightCycler® RNA Master HybProbe
Easy-to-use reaction mix for one-step RT-PCR using the LightCycler® Carousel-Based System.
For general laboratory use.
Easy-to-use Reaction Mix for One-Step RT-PCR, using HybProbe probes with the LightCycler® Carousel-Based System.
- The kit is particularly suitable for difficult RNA populations since the elevated incubation temperature during the RT step will help to overcome secondary structures, and the hot start feature will minimize mispriming during the RT step.
- The LightCycler® RNA Master HybProbe provides convenience, high performance, reproducibility, and minimized contamination risk.
- LightCycler® RNA Master HybProbe, 2.7x concentrated
- Mn(OAc)2 Stock Solution, 50 mM
- Water, PCR Grade
The LightCycler® RNA Master HybProbe is a easy-to-use hot start reaction mix, specifically adapted for one-step RT-PCR in LightCycler® Capillaries using the LightCycler® Carousel-Based System Instruments and HybProbe probes as the detection format.
Note: LightCycler® RNA Master HybProbe can be used in conjunction with Uracil DNA Glycosylase, heat-labile for carryover prevention during PCR.
The hot start feature is achieved by using Tth DNA Polymerase in combination with Aptamers.
Tth DNA Polymerase is a thermostable enzyme with RNA-dependent Reverse Transcriptase activity and DNA-dependent Polymerase activity, allowing the combination of RT and PCR in a single-tube reaction.
Aptamers are dedicated oligonucleotides that bind in the active center of the polymerase and prevent attachment to nucleic acid targets at temperatures below the optimal reaction temperature of the Tth enzyme. Therefore, no primer elongation occurs during setup of the reaction and the following heating phase prior to the RT step. At higher temperatures, the Aptamers are released from the enzyme, and RT or DNA polymerization can be initiated. In addition, the recommended incubation temperature for Reverse Transcription with Tth (+61°C) is helpful to overcome secondary structures of RNA. This results in highly specific and efficient cDNA synthesis, which leads to highly specific and sensitive PCR.
Function test: The LightCycler® RNA Master HybProbe is function tested using the LightCycler® Control Kit RNA, according to the kit protocols.