Technology

• Hairpin architecture: Stem holds fluorophore and quencher
  in proximity; loop is complementary to the target.
• Signal-on binding: Target binding opens the stem–loop,
  separating dye and quencher, restoring fluorescence.
• Reversible detection: Beacon remains intact (non-cleaving), enabling melt analysis and reuse across cycles.
• Thermal tuning: LNA/BNA or 2′-modifications raise T_m for
  shorter, more specific beacons.
• Multiplex-ready: Distinct dyes with minimal spectral overlap
  support multi-target panels.

Benefits

• Ultra-low background when unbound → clean baselines.
• Single-nucleotide discrimination from stem-loop energetics.
• Isothermal compatibility (LAMP/RPA) and real-time qPCR/dPCR.
• Non-destructive (no probe cleavage), enabling post-run melt analysis.
• Reproducible manufacturing with HPLC + MS QC.

We tune stem strength, loop length, and backbone chemistry to your temperature and matrix.

• Loop (target region): ~18–30 nt (miRNA: 15–22 nt with LNA/BNA).
• Stem: 5–7 bp, GC-rich; avoid complementarity within the loop.
• T_m rules: T_m(stem) ≈ assay temp + 5–10 °C; T_m(loop–target) ≥ assay temp + 7–10 °C.

• Placement: Avoid terminal 5′-G (can quench some dyes); minimize secondary structure in the target.
• Concentration: Start 100–250 nM (optimize 50–400 nM).
• Chemistry: Consider 2′-OMe/2′-F or LNA/BNA mix for short targets and SNPs.

• Multiplexing: Separate dye emissions by ≥40–60 nm; balance brightness.
• Controls: NTC and mismatch controls; verify closing/opening by melt.
• Imaging: Add PEG/AEEA spacers to reduce self-quenching near dyes.