This product is a comprehensive one-step lyophilization-compatible amplification system that includes: reaction buffer containing Mg²⁺, dNTPs, and other essential components; RT enzyme mix composed of Bst DNA polymerase and a heat-resistant reverse transcriptase; fluorescent dye for real-time detection; and lyophilized protectant for optional freeze-drying preparation. Users can simply mix the reaction buffer, RT enzyme mix, fluorescent dye, and primers, then add the template to start the reaction. If needed, the lyophilized protectant can be added to create a system suitable for freeze-drying.

1. Thaw buffer at room temperature, invert several times, centrifuge.
2. Two preparation types:
2.1 Liquid Mix:
Loop-mediated Amplification Buffer: 12.5 μL
25× Dye: 2 μL
RT-Enzymes Mix: 2.55 μL
10× Primer Mix: 5 μL
Target DNA/RNA: x μL
Nuclease-free water: up to 50 μL
2.2 Freeze-drying Mix:
Buffer: 12.5 μL, Lyophilized protectant: 20 μL, Dye: 2 μL, RT-Enzymes Mix: 2.55 μL, Water: up to 50 μL
Freeze-dry in 50 μL system
Reconstitute: add 10× Primer Mix: 5 μL, Target DNA/RNA: x μL, Water: up to 50 μL
Notes: 10× Primer Mix includes 16 μM FIP/BIP, 2 μM F3/B3, 4 μM Loop F/B. Use DEPC water for template dissolution.
3. Run at 65°C for 60 s, 30 cycles on PCR instrument (e.g., ABI7500).

Salt may accumulate in buffer; mix thoroughly at room temperature.
Optimize temperature between 62°C-68°C based on primer conditions.
Standardize processes: reaction prep, lyophilization, sample handling.
Prepare system in ultra-clean bench. Add templates in fume hood to avoid contamination.

Ship with dry ice packs. Store at -20°C. Avoid repeated freeze-thaw cycles. Stable for 12 months under recommended conditions.