LightCycler® 480 RNA Master Hydrolysis Probes
Easy-to-use hot start reaction mix for one-step RT-PCR using the LightCycler® 480 and 96 Instruments, and the LightCycler® Nano Instrument.
For life science research only. Not for use in diagnostic procedures.
The kit provides reagents, including an RNA master mix (with buffer, nucleotides, and enzyme), a Mn(OAc)2 stock solution, PCR-grade water, and enhancer solution. The LightCycler® 480 RNA Master Hydrolysis Probes can be used in conjunction with Uracil DNA Glycosylase, heat-labile for carryover prevention during PCR.
- Significantly reduce assay time to less than 45 minutes.
- Increase signal dynamics and sensitivity (down to 0.1 pg of total RNA) by using the kit's special enhancer solution.
- Perform multiplex detection of target and reference genes.
- Combine analysis of DNA and RNA viruses in one reaction.
- LightCycler® RNA Master Hydrolysis Probes, 2.7x concentrated
- Mn(OAc)2 Stock Solution, 50 mM
- Water, PCR Grade
- Enhancer solution, 20x concentrated
The LightCycler® 480 RNA Master Hydrolysis Probes is an easy-to-use hot start reaction mix, specifically adapted for one-step RT-PCR under the rapid and accurate cycling conditions of the plate-based LightCycler® 480 and 96 Instruments, as well as the LightCycler® Nano Instrument using hydrolysis probes (e.g., Universal ProbeLibrary probes) as detection format.
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 reverse transcription (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 RT 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.
Hot start with Aptamers is highly effective and very convenient because it does not require additional incubation steps, pipetting steps, or an extension of reaction time. The hot start protocol with Aptamers does not interfere with other enzymatic processes, the online detection of amplification products, or subsequent handling steps.
Function test: The LightCycler® 480 RNA Master Hydrolysis Probes is function tested on the LightCycler® 480 Instrument using beta-actin as a target gene, HeLa total RNA, and a Universal ProbeLibrary probe.