# Rice seed germination
All ddH2O used for this process should be sterilized.
## **Step 1: Homogenization**
1. Dehusked the seeds. Select only seeds that are well-developed, fully-filled, clean, and transparent.
2. Dry-incubate the seeds in dark under 45°C for 24 hours.
## **Step 2: Sterilization**
1. Soak the seeds in 3% bleach solution ( add one drop of detergent ) and gently inverting / shaking for 20 mins.
2. Wash at least 6 times with ddH2O, until the bubble has gone and water is clear.
## **Step 3: Priming and imbibition**
1. Remove the water thoroughly with pipette.
2. (optional) Seed priming. Soak the seeds in 3% KNO3 solution under room temperature for 24 hours;
> - The KNO3 priming method is not effective in our case
3. Soak the seeds in 1 ~ 2% Plant Preservative Mixture™ ( PPM™ ) solution and incubate at 30°C in dark for 16 ~ 24 hours.
> - For example, add 300 µL PPM into 30 mL sterilized ddH2O.
## **Step 4: Sowing**
1. Remove the PPM solution.
2. Sow the seeds on suitable medium.
> - The seed must be fully embedded into the medium, otherwise, the seedling will push itself up during root penetrating into the gel.
> - Optional: Add 0.05 ~ 0.2% ( v / v ) PPM™ to the medium, if it is difficult to control contamination. The PPM™ can be autoclaved together with the media.
Done !!!