Lyo-ready Enzymes for Molecular Diagnostics

Glycerol-free vs lyo-ready: what changes in the tube

The standard liquid form of each enzyme contains glycerol, which keeps it stable at −20 °C but never leaves the product during drying: it stays behind as a syrupy residue that prevents a dry, porous cake from forming. Each enzyme on this page is therefore also supplied in a glycerol-free identity with an F-prefixed catalog number (FE03 for E03, for example), so it can go into a reaction that will be freeze-dried.

Glycerol-free is what the enzyme itself has to be. Whether a complete reaction dries well is decided by the whole mix: buffer, primers and probes, lyoprotectant, fill volume and drying cycle. Disaccharides such as trehalose and sucrose form a glassy matrix around the enzyme and replace the hydrogen bonds that water provided, so the protein keeps its native fold in the dry state. For complete reactions that are ready to dry, see our lyo-ready master mixes.

Build a freeze-dried assay from individual enzymes

1. Choose the enzymes. A hot-start Taq or Tth for PCR and qPCR, or a Bst for LAMP; a reverse transcriptase for RNA targets; UNG/UDG for carry-over control; RNase H II for probe-based LAMP. Order each in its glycerol-free identity.
2. Add a lyoprotectant. AS212-17 for DNA systems or AS212-156-5 for RNA systems. Both are 2× concentrates: 12.5 µL in a 25 µL reaction gives 1×. AS412-9 is a 4× concentrate (6.25 µL in 25 µL). The water volume can be trimmed to improve the dried form. All three are sold separately.
3. Freeze-dry in the final format. Beads, 8-tube strips, 96-well plates, pre-filled tubes or vials.
4. Compare with the liquid control. Run the dried reaction against the same-batch liquid reaction, before and after storage, as in our own stability studies.

If you would rather not develop the drying cycle in-house, our lyophilization service covers cycle development, pilot lots and production, and the feasibility assessment of your formulation is free of charge.

Performance after lyophilization

Mix-level data. The results below were generated with two complete BIORI lyo-ready master mixes, primers and probes included, not with the individual enzymes on this page.

• Real-time: FM2181-4 (4-plex qPCR, DNA) and FM5144 (4-plex RT-qPCR, RNA) stored 18 months at room temperature, every channel within ΔCt ≤ 1 and ΔRn within 10% of the same-batch liquid control.
• Accelerated: 37 °C for 150 days and 45–55 °C for 12–90 days across 8-tube strips, vials and beads.
• Shipping stress: beads left open 4 hours at 20–25% RH, sealed, then held 30 days at 55 °C; amplification stayed consistent with the liquid control.

See the full stability data on the lyophilization service page.

Lyophilization or air-drying?

Freeze-drying removes water from a frozen reaction under vacuum; air-drying removes it by evaporation, without freezing. Our stability data cover freeze-dried formats only.