Question 1 :【How to handle the plasmid】 

A: The item is lyophilized form,  Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid. 

Question 2:【Plasmid transformation steps---E.coli】 

A: Operation steps:

The item is lyophilized form,  Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.

Transformation steps：

1)Take corresponding competent cells out from the refrigerator at – 80℃, thawed on the ice box at room temperature then labeled it;

2) Add 2μl plasmid into the competent cells and placed at 4℃ for 30 min;

3. Heat shock in a 42℃ water bath for 1 min, and then place on the ice box for 2 min immediately ;

4. Added 900μl LB medium in each tube, shaking at 37℃, 150 rpm × 45 min;

5. Add 100 μl conversion solution into the corresponding resistant plate, pour the glass beads, and spread the liquid evenly; [2]

6. Invert the plate in constant temperature incubator at 37℃ for overnight.

*Cautions:
[1] The sterile centrifugal tube can be used in the sensory state.
[2] According to your request-Step 5, Centrifuge the conversion solution at 6000rpm for 3min, then leave 100 μ l of supernatant, mix and precipitate, and add all into the corresponding plate.
[3]Transformation should be accomplised in the ultra clean worktable.