Short probes. High Tm. Real discrimination. Minor Groove Binder (MGB) hydrolysis probes engineered for qPCR and SNP genotyping with lower background and cleaner ΔRn.
MGB probes incorporate a minor‑groove‑binding moiety at the 3′ end alongside a non‑fluorescent dark quencher. The MGB raises duplex stability so you can use shorter probes (≈14–18 nt) without losing Tm, tightening SNP discrimination and reducing leak.
Standard layout: 5′ Reporter — probe — 3′ MGB‑NFQ (3′ blocked). Multiplex with spectrally separated reporters.
Optional: Endotoxin (LAL), bioburden, sterility, ds%/UPLC, functional qPCR.
MGB enables shorter probes; keep amplicons 70–150 bp for crisp kinetics. Maintain probe Tm ~10 °C above anneal/extend to minimize breathing.
Map dyes to non‑overlapping channels, verify your instrument’s color compensation, and balance probe concentrations per channel. We can deliver a pre‑balanced 2–4‑plex kit on request.
Hydrolysis probes without MGB for standard 5′‑nuclease assays.
Locked nucleic acid designs for added mismatch discrimination.
Validated primer design and synthesis matched to your MGB probe.
Both boost Tm and mismatch discrimination. MGB stabilizes via minor‑groove binding at the 3′ end, enabling short probes with a dark quencher. LNA increases Tm through locked sugar chemistry across the sequence. Choose MGB for compact hydrolysis probes; choose LNA when you need localized Tm boosts within the span.
Avoid 5′‑G adjacent to the dye; keep GC 40–60%; center the SNP; avoid >3 identical bases; check for hairpins/duplexes (ΔG < −2 kcal/mol); maintain probe Tm 6–10 °C above primer Tm; keep amplicon 70–150 bp.
Map dyes to well‑separated channels (e.g., FAM/HEX/Cy3/ROX/Cy5), verify color compensation on your instrument, and titrate each probe for balanced ΔRn. We can deliver pre‑balanced mixes.
Any 5′‑nuclease‑competent real‑time PCR mix (Taq‑based) works. Hot‑start Taq mixes with ROX reference are fine; ensure your reporter is not reserved as a passive reference on your platform.
Use slightly longer probes (17–20 nt) or shift to a reporter with strong signal (e.g., FAM/Cy3). Optimize Mg²⁺ and anneal temperature; consider DMSO/Betain per platform guidelines.
Dual‑HPLC, UPLC purity %, ds‑content, endotoxin (LAL), bioburden, sterility, and functional qPCR on request.
Spin down, reconstitute in nuclease‑free water or TE to 100 µM, aliquot to avoid repeated freeze–thaws, store −20 °C protected from light. Working dilutions (e.g., 10 µM) should be kept at 4 °C for <2 weeks.
Yes—96‑well or 384‑well plates with custom layout and barcodes; include your pick list with desired concentrations/volumes.
Often yes—we support common FAM/HEX/Cy3/ROX/Cy5 families and MGB‑class dark quenchers. Provide your current catalog code or excitation/emission requirements.
Target sequence or accession with ±100 bp context, platform/instrument, desired reporter channels, and any constraints (GC window, SNP position, amplicon length).
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