Cat. No. JY0808
Product Overview
RPApex Fluorescence/LFS uses enhanced RAA technology for rapid isothermal amplification at 39–42℃ with integrated exonuclease/Nfo nuclease for fluorescence or lateral flow strip detection.
Principle
- Recombinase-primer complex invades target DNA
- SSB proteins maintain open-chain state
- DNA polymerase initiates strand synthesis
- Exponential amplification in 10–20 minutes
- Contains exonuclease/Nfo nuclease for fluorescence or LFS detection
- Ready-to-use lyophilized pellets
Primer Design Guidelines
- Length: 28–35 nt (longer than PCR primers)
- Too short (<28 nt): reduced efficiency
- Too long (>35 nt): secondary structures interfere
- Concentration: Optimize via gradient (0.2–0.6 μM)
Probe Design
- Length: 28–35 nt, complementary to target
- Introduce THF (dSpacer) for enhanced cleavage
- Fluorescence: 5′ fluorophore (FAM), 3′ quencher (BHQ1)
- LFS: 5′ antigen tag (FAM), 3′ blocking group (C3-spacer)
- Avoid secondary structures; verify with Primer Premier or OligoAnalyzer
Amplicon Length
- Recommended: 80–300 bp
- Too long (>300 bp): reduced efficiency
- GC content: 40–60% preferred
Storage and Stability
- Unopened: Store at 25–30℃, dry and light-protected. Shelf life: 12 months
- After opening: Tighten cap, continue storage at 25–30℃, dry and light-protected. Shelf life: 2 months