LightCycler® RNA Master SYBR Green I
Easy-to-use reaction mix for one-step RT-PCR using the LightCycler® Carousel-Based System.
For life science research only. Not for use in diagnostic procedures.
The LightCycler® RNA Master SYBR Green I provides reagents, including RNA Master Mix (with buffer, SYBR Green I, nucleotides, and enzyme), a Mn(OAc)2 stock solution, and PCR-grade Water. The kit is specially designed for use with the LightCycler® 2.0 System Instruments. The LightCycler® RNA Master SYBR Green I can be used in conjunction with heat-labile Uracil DNA Glycosylase for carryover prevention during PCR.
- The kit is particularly suitable for difficult RNA populations because 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 SYBR Green I provides convenience, high performance, reproducibility, and minimized contamination risk. Only template RNA, primers, and additional Mn(OAc)2 (if necessary) must be added.
- LightCycler® RNA Master SYBR Green I, 2.7x concentrated
- Mn(OAc)2 Stock Solution, 50 mM
- Water, PCR Grade
The LightCycler® RNA Master SYBR Green I is an 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 SYBR Green I dye as the detection format.
Note: LightCycler® RNA Master SYBR Green I 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 with 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 in overcoming 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 SYBR Green I is function tested using the LightCycler® Control Kit RNA, according to the kit protocols.