This product contains reaction buffer, RT enzymes mix of Bst DNA polymerase and heat-resistant reverse transcriptase, lyophilized protectant and fluorescent dye components. The reaction buffer contains Mg 2+, dNTP and other necessary components for amplification, users can directly mix the reaction buffer, RT enzymes mix, fluorescent dye, primer, and then add the template. If necessary, users can add the lyophilized protectant to prepare lyophilizable system.
Transportation and storage
Transported with dry ice, stored at -25～-15℃. Avoid repeated freezing and thawing, the product is valid for 12 months.
- Thaw the reaction buffer to be used at room temperature. Vortex briefly or invert tubes several times to mix thoroughly, then centrifuge
to collect the liquid to the bottom of the tube.
- Prepare reaction mix as described below
10×Primer Mix: FIP/BIP 16μM，LoopF/B 4 μM，F3/B3 2 μM. Probe concentration may be optimized according to specific situation.
1.3. Reaction program
*Collect fluorescent signal.
2 Using fluorescent dye
1.1 Thaw buffer to be used at room temperature. Vortex briefly or invert tubes several times to mix thoroughly. Centrifuge to collect material and place on ice.
1.2. Prepare reaction mix as described below
1) Salt may appear in the bottom of the buffer tube, vortex briefly or invert tubes several times to mix thoroughly at room temperature.
2) The reaction temperature can be optimized between 62°C and 68°C according to the primer conditions.
3) The experiment shall be conducted in a standardized manner, including the preparation of reaction system, sample treatment and sampleaddition. It is also suggested to prepare reaction system in the ultra clean table and add templates in the fume hood of other rooms to avoid false
For DNA or RNA isothermal amplification.