KM71 感受态细胞(化转)/ KM71 chemically Yeast Express Competent Cells

KM71 Chemically Competent Cell

Packing 100 x 0.1 ml/tube

Store at –80°C

Genotype

 arg4 his4 aox1::ARG4

Alternative name
KM71 chemically Competent Cells,KM71 Competent Yeast,KM71 Competent Cells, KM71 chemically Yeast Express Competent Cells

Transformation Protocol

1.Please store KM71 Chemically Competent Cell in -80°C for late use.

2. Thaw a tube of KM71 Chemically Competent Cell锛?00μl 锛塷n ice.

3. Heat shock Carrier DNA at 95-100°C then immediately transfer the tube back into ice for 5min,2times.

4. Add 2-5µl plasmid DNA, Carrier DNA 10ul and PEG/LiAc 500ul to the tube. Carefully flick the tube 4–5 times to mix cells and DNA.

5.Place the mixture in the water 30°C for 15 minutes. (Flip it 6-8times at 7.5min)

6. Place the EP Tube in the water 42°C for 15 minutes. (Flip it 6-8times at 7.5min)

7. Pellet the mixture by centrifugation at 500rpm/min for 40s. Pour off cell supernatant, then resuspend the cell pellet in the ddH2O 400ul..

8. Pellet the mixture by centrifugation for 30sec., Pour off cell supernatant. resuspend the cell pellet in the ddH2O 400ul.

9.Spread the cell on the agar plates by antibiotic, then place it on 29°C for 48-96hr.

Caution:

1.STORAGE AND HANDLING: Competent cells should be stored at

–80°C. Storage at –20°C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above –80°C, even if they do not thaw.

2. This product is FOR RESEARCH USE ONLY!