RTGreen is a dye for quantitative real-time PCR (qPCR). It is a novel DNA-binding dye specially designed for qPCR and high-resolution melting (HRM) analysis. Via an "on-demand release" mechanism, it greatly reduces inhibition of PCR amplification. RTGreen outperforms SYBR Green I in many aspects. While possessing similar spectral properties, RTGreen differs from SYBR Green I with three key characteristics:
1. It exhibits much lower PCR inhibition than SYBR Green I, enabling qPCR experiments with fast PCR protocols.
2. It can be applied at higher concentrations in assays to generate significantly stronger amplification signals than SYBR Green I. Higher concentrations of RTGreen eliminate the "dye redistribution" artifact, making RTGreen suitable for both multiplex PCR and high-resolution melting (HRM) analysis.
3. This assay is increasingly applied for post-PCR genotyping and heteroduplex analysis.
Another advantage of RTGreen is its safety. Conventional DNA-binding dyes carry potential mutagenicity. With this concern in mind, we developed RTGreen. It cannot penetrate cell membranes and therefore cannot bind to genomic DNA in live cells, guaranteeing its safety. In contrast, all other commercial fluorescent PCR dyes can enter cells within minutes. For instance, SYBR Green I is a well-known mutagen with greater environmental toxicity than ethidium bromide (EB).
Product Features:
1. Ultra-high sensitivity: Delivers maximum PCR amplification signal when used at the recommended concentration.
2. Minimal PCR inhibition: The "on-demand release" mode of dye-DNA binding results in far less PCR inhibition by RTGreen compared with SYBR Green I.
3. Compatible with fast PCR: Minimal interference with PCR, which greatly shortens PCR extension time.
4. Ideal for HRM analysis: Free from the "dye redistribution" artifact, compatible with post-PCR high-resolution melting (HRM) analysis.
5. Compatible with multiplex PCR: No dye migration among amplicons at the recommended working concentration.
6. Excellent stability: Highly stable under most biochemical conditions, storable at room temperature and tolerant to repeated freeze-thaw cycles.
7. High safety: Cell membrane penetration assays demonstrate that RTGreen barely crosses cell membranes.
8. Excellent compatibility: Similar spectrum to SYBR Green I, compatible with most qPCR instruments. No modifications to your existing protocols or equipment are required.
Recommended PCR System:
Suggested PCR reaction system (50 μL): Optimize according to different primers and templates. The system below is designed for non-hot-start Taq polymerase, with 1× RTGreen recommended.
| Reagent | Final concentration in the mixture | | 5μL each of 2mM dNTP | 0.2mM each | | ddH₂O to a final volume of 50μL | Adjust to final volume | | 5μL of 10× polymerase buffer without magnesium | 1× | | 0.1-1μM each of primers (final concentrations) | 0.1-1μM | | 1-5 units of Taq DNA polymerase | 1-5 units per reaction | | 2.5μL of 50mM MgCl₂ | 2.5 mM | | 2.5μL of 20× RTGreen | 1× |
|
ZJ26F0939511