In vitro culture experiment of carrot root

In vitro culture experiment of carrot root

[ Plastic Petri Dish,Petri Dish 60mm,90mm Petri Dish ]

1. Rinse the carrots with tap water, remove 1-2mm of the skin with a paring knife, and cut into slices about 10-40mm thick. The following steps are all performed aseptically.
2. After the carrot slices are treated with 70% ethanol for 30-60 seconds, rinse with sterile water, then soak in 0.1% mercuric chloride or saturated bleach solution for 10 minutes, and rinse with sterile water for 3-5 times.
3. Place the carrot slices in a petri dish, fix the carrot slices with tweezers in one hand, and punch holes parallel to the vertical axis of the tissue slices with the other hand. Each hole should be made close to the vascular cambium, making sure to penetrate the tissue. The punch was then withdrawn from the tissue piece, and the cylinder was gently pushed out of the punch with a glass rod and placed in a petri dish of sterile water. Repeat the punching step until a sufficient number of tissue cylinders are prepared.
4. Take out 1-2 carrot cylinders with tweezers, put them into another petri dish containing sterile water, and use a razor blade to cut off 2mm long tissue at each end of the cylinder. Cut the remaining tissue into small discs about 2mm thick. The forceps and scalpel are flame sterilized several times throughout the cutting operation and cooled before use.
5. Transfer the disc to sterilized filter paper (one piece at a time) with tweezers, blot the water on both sides of the disc, and immediately inoculate the surface of the medium. Inoculate 6-8 tablets per bottle.
Note: The triangular flask is inclined to a certain degree during inoculation, and the whole inoculation process, such as holding tweezers by hand, burning, etc., should not be completed directly above the medium to reduce the chance of contamination.
6. Incubate the culture in a 25°C incubator. One part can be cultivated in the light, and the other part can be cultivated in the dark, and the response of the induced callus in the light and the dark place can be compared.

[ Plastic Petri Dish,Petri Dish 60mm,90mm Petri Dish ]