One-Step TaqProbe qRT-PCR-iCycler

Details for Product No. ABIN4219232,
Quantitative real-time PCR (qPCR)
Purpose Convenient real-time RNA quantification in one EASY step.
Specificity - Streamlined protocol in a simple, single-tube reaction set-up
- High-quality, full-length cDNA from as little as 0.01pg of RNA template
- Fully optimized for detection of low-copy genes
- Simple set-up for any RNA template (e.g. mRNA and total RNA)
- Reduces pipetting steps to minimize the risk of contamination and error.
Characteristics One-Step RT-PCR Kit contains all necessary reagents for both reverse transcription and PCR amplification in a single reaction tube. Specifically, the One-Step RT-PCR Kit combines EasyScriptTM RTase and BestaqTM DNA Polymerase in a convenient format for highly sensitive and specific RT-PCR with any RNA template. Our proprietary RT-PCR Buffer contains stabilizers and enhancers that optimize the two reactions into a "single step". Together with our specially formulated RT-PCR Buffer, this One-Step RT-PCR Kit offers endusers an efficient, easy to use, and reliable alternative to conventional "twostep" sequential RT-PCR.

One-Step TaqProbe qRT-PCR Kit uses a combination of high-quality enzymes in a proprietary buffer system to deliver precise and accurate sample analysis for high-throughput applications. This kit offers ultimate convenience in addition to consistent performance in terms of high sensitivity and superb signal-to noise ratio.
While a one-step/single tube setup provides overall convenience and reduces room for error, abm's One-Step TaqProbe qRT-PCR Kit offers additional advantages:
- Improved fidelity and yield for reverse transcription
- Prevention of template (RNA) degradation with RNaseOFF Ribonuclease Inhibitor.
- Superb performance with respect to sensitivity and signal-to-noise ratio.
- Significant reduction in non-specific PCR amplification by utilizing HotStart Taq DNA polymerase in the enzyme mix.
Components This kit includes 50X qRT-PCR Enzyme Mix and TaqProbe qPCR MasterMix
Application Notes - Gene-expression analysis
- Transcription analysis
- Gene cloning
- High throughput applications
- Virus detection and quantification

100 rxn (20 μl/rxn)
1. Aliquot reagents to avoid risk of contamination and repeated freeze-thaw cycles.
2. Avoid high primer concentrations, a concentration of 100 nM to 300 nM of each primer usually gives the best results.
3. Probe concentration may require optimization, typically a 3:1 to 1:1 primer:probe ratio will give good amplification.
4. A very effective way to get tight Ct among replicates is to reduce pipetting error, this can be achieved by: 1) preparing amplicon specific pre-mix, 2) using repeating pipettes and 3) keeping the pipetting volume within the manufacturer's suggested range.
5. For optimal results, it is recommended that the primers are 18-22 nucleotides in length with a Tm of 58°C-60°C and the size of target is about 100-250 bp.

Restrictions For Research Use only
Storage -20 °C
Storage Comment Stored all components at -20°C. Protect the TaqProbe qPCR MasterMix from light.