This product contains a reaction buffer, RT-Enzyme Mix (comprising Bst DNA polymerase and heat-resistant reverse transcriptase), lyophilized protectants, and chromogenic dye components for visual detection. To prepare the reaction system, simply mix the reaction buffer, enzyme mix, and primers, then add the template. If lyophilization is required, lyophilized protectants can be added directly, and the complete mixture can be freeze-dried using a lyophilizer. During use, only primers and templates need to be added to the lyophilized reagents. This kit offers a reliable and efficient method for visual nucleic acid amplification detection, which negative reaction is indicated in red and positive reaction is indicated by a change to yellow, ideal for point-of-care or resource-limited settings.

1. Thaw buffer at room temperature, invert to mix, centrifuge.
2. Choose system type:
2.1 Liquid Mix: Buffer (with dye): 10 μL, RT-Enzymes Mix: 2.8 μL, 10× Primer Mix: 5 μL, Template DNA/RNA: x μL, Water: up to 50 μL
2.2 Freeze-drying Mix: Buffer (with dye): 10 μL, Lyophilized protectant: 20 μL, RT-Enzymes Mix: 2.8 μL, Water: up to 50 μL. Freeze dry in 50 μL system. Rehydrate with 10× Primer Mix: 5 μL, Template DNA/RNA: x μL, Water: up to 50 μL.
Primer Mix includes 16 μM FIP/BIP, 2 μM F3/B3, 4 μM Loop F/B. Use DEPC-treated water for nucleic acids.
3. Incubate at 65°C for 30–45 minutes. Yellow indicates positive, red indicates negative (visible to the eye).

Salt may accumulate in buffer—vortex or invert before use. Optimize temperature between 62°C–68°C. Avoid prolonged exposure to air. Do not use Tris-based buffers—use ddH2O. Standardize reaction and lyophilization steps. Prepare mixes in ultra-clean bench and handle templates in fume hood to prevent contamination.

Ship with dry ice. Store at -25 to -15°C. Avoid repeated freeze-thaw cycles. Stable for 12 months under recommended conditions.