Cell clone
Soft agar clone: u200b u200bclonal formation rate reflects two important chaos of cell population dependence and proliferation (123] 1. Get samples, cultured cells,
2. Adjust cell suspension The density is 1 x 103 / ml cells.
3. Preparation of underlying agar, completely dissolved 5% agar and 37 * c left and right pre-temperature fresh fully medium in 1: 9 ratio in 40 ^ c, adding a culture dish (straight
In 60 mm), the doni was 0.5% agar medium 2 mL, and the agar was completely solidified at room temperature.
4. Preparation of the upper agar, 37C density 10,000 cell suspension 1.5m is transferred into the small beaker, adding 40c, 5% agar, etc., is 0.25% semi-solid
grease culture base. The mixed semi-solid agar medium immediately added a petri dish on the underlying agar, and agar solidified at room temperature. 37C, 5% CO2 stationary culture 2-
3 weeks.
5. When there is a cloning in the culture dish, it is visible, and the culture is terminated, discarded, and the PBS is carefully dipped twice, and the air is dried. Methanol was fixed for 15 minutes, and after abandoning methanol
air dried. It was stained with GIEMSA dye for 10 minutes, and the water was slowly washed away, and the air was dried.
Experiment Period and Delivery Standard
1. Experimental Period: 1-2 Week
1. Dilution laying method
2. Breeding layer cloning
3. Capillary cloning method
4. Fluorescence activation sorting method