One-Step EvaGreen qRT-PCR-No Dye

Details for Product No. ABIN4219226,
Quantitative real-time PCR (qPCR)
Purpose Convenient real-time RNA quantification in one-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
- Fully optimized for detection of low-copy genes
- Simple set-up for any RNA template
- Reduces pipetting steps to minimize the risk of contamination
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 EvaGreen qRT-PCR Kit uses a combination of high-quality enzymes in a proprietary buffer system to deliver precise and accurate sample analysis in a high-throughput format. This kit offers ultimate convenience to the end-user in addition to guaranteed performance with respect to high sensitivity, superb signal-to noise ratio, and complete elimination of primer dimmers. The One-Step EvaGreen qRT-PCR Kit utilizes a mutated recombinant M-MuLV Reverse Transcriptase which exhibits limited RNase H activity. RNaseOFF Ribonuclease Inhibitor and a proprietary RTase additive are blended into the RTase's storage buffer to protect RNA template from degradation and to improve the fidelity during reverse transcription. The chemically modified Hotstart Taq polymerase, included in our MasterMix, significantly reduces non-specific PCR amplification observed with regular Taq polymerase. The user-friendly one-step/single tube setup minimizes the chance of introducing human-error and contamination. Overall, abm's One-Step EvaGreen qRT-PCR Kit demonstrates market-leading sensitivity, efficiency, and reliability.
Components This kit includes 50X qRT-PCR Enzyme Mix and EvaGreen 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 contamination and to avoid repeated freeze-thaw cycles.
2. Primer concentration should not be high, a concentration of 100 nM to 300 nM of each primer usually gives the best results.
3. A very effective way to get tight Ct among replicates is to reduce pipetting error, this can be achieved by: preparing amplicon specific pre-mix, using repeating pipet, and keeping pipetting volume in manufacture suggested range.

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