The document is an operation manual for GMO Hunter MON810, which describes the operation steps and composition of the fluorescence PCR reaction and positive control.
The MON810 GMO Hunter kit includes a real‑time PCR master mix, positive control, and reagents that must be stored at –20 °C and protected from light, e.g., wrapped in foil. Each PCR reaction uses a total volume of 20 µl with 11.2 µl master mix, 8.8 µl water, and 100‑200 ng DNA template.
The procedure describes thawing reagents on ice, preparing reaction mixes, loading PCR tubes into a LightCycler® Real‑Time PCR system, and running 40 cycles (95 °C 15 s, 60 °C 60 s) with fluorescence data collection. It also explains data interpretation, the use of positive/negative and spiking controls, and provides technical support contact information. This manual is written in Chinese.
| DNA template amount | 100‑200 ng |
|---|---|
| Positive control volume | 2 µl |
Run 1 initial cycle at 95 °C for 2 min, then 40 cycles of 95 °C for 15 s and 60 °C for 60 s, collecting fluorescence at the FAM 530 nm channel.
Each test sample should be run in two PCR replicates, with a positive control (2 µl) and a negative control (2 µl water).
A spiking control (adding 1 µl positive control to the sample) verifies that PCR inhibition is not causing a false‑negative result.