One.Step RT-qPCR Probe Kit with ROX (2X concentrated)
Opis
Features:
- The kit contains all reagents required for RT-qPCR in a set to ensure fast and easy preparation with a minimum of pipetting steps. Just add template, primers and the dual labeled fluorescent probe.
- The One.Step RT-PCR Kit is highly sensitive: less than 10 pg to 100 ng total RNA or < 1 pg to 20 ng poly(A) RNA (mRNA) can be detected when using highly expressed transcripts.
Description:
The enzyme mix is based on a new formulated reverse transcriptase, a Hot-Start Polymerase and RNase Inhibitor. The set is providing increased specificity, high cDNA yield and improved efficiency for highly structured and long cDNA fragments. The reaction buffer includes extrapure dNTPs; ROX and reaction enhancers. The basis is a Real-time PCR with dual labeled probes and multiplexing capacity.
Principal:
In the RT-step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. The hot-start polymerase activity is blocked.
In the first cycle of the PCR step the Hot-Start DNA polymerase activity is switched on and it synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template. The kit is optimized for all real-time PCR cyclers who are compatible with the evaluation of the ROX reference signal.
The One.step Realtime RT-PCR Master Mix from GeneON offers a tremendous convenience when applied to analysis of targets from multiple samples of RNA and minimizes the risk of contaminations.
Platforms: The Kit is suitable for all block-based Thermocycler. Stringent Quality Tests on ABI StepOne plus PCR Cycler
Components of Maximo.OneStep RT-qPCR Master Mix (for probes):
- Enzyme-mix: HotStart Taq Polymerase, Reverse Transcriptase, RNase Inhibitor and enhancers, Reaction buffer containing extra pure dNTPs and ROX
- RNase-free water
Protocols / Manuals
The datasheet includes detailed protocols for:
1.) RT-PCR assay without sample denaturation (as a standard RNA/primer combinations)
2.) RT-PCR assay with sample denaturation (RNA/primer with a high degree of secondary structure)
2.a) Preparation of RNA / Primer Mix
2.b) Denaturation and primer annealing
2.c) Preparation of the RT-PCR Mix
3.) Reverse Transcription and thermal cycling
Storage: @ -20°C, avoid frequent thawing and freezing, store all components with ROX in the dark
For optimal results it is recommended to make an individually optimization for each RNA / primer pair.
Manufactured and quality-controlled in accordance with ISO 9001:2000.
Dane techniczne
Opakowanie | 100 x 25 μl rxns |