PCAG Cre: GFP|357422 |BNCC

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pCAG-Cre:GFP

Subculture procedure	1. Dissolution: After receiving the plasmid powder, please add 20 μ l of sterile water to the bottom of the tube, dissolve the plasmid, and let it stand at room temperature for 1 minute; 2. Mixing (adsorption plasmid): Mix 200 μ l of competent cells and 5-10 µ l of plasmid DNA, and place on ice for 30 minutes; 3. Heat shock introduction: let stand at 42 ℃ for 90 seconds; 4. Shrinkage film pores: ice bath for 2 minutes; 5. Repair culture: Add 800 µ l LB liquid medium to each tube and incubate at 37 ℃ for 1 hour at 150 r/min; 6. Screening and cultivation: Apply an appropriate volume (100 µ l) of revived cells onto corresponding resistant LB plates, place them upright in the plate for 30 minutes (the agar surface must be dry), invert and culture for 12-16 hours, and colonies will appear. 7. Extraction: Select monoclonal colonies into the corresponding resistant LB liquid medium, shake and culture for 12-16 hours, and extract plasmids according to experimental needs.
Growth conditions	LB+ampicillin, 37 ℃ (cloned strain DH5 α, 6563bp)
Storage conditions	2-8 ℃
Safety level	1
Sharing mode	Public welfare sharing

Product format: freeze dried, 1.0-2.0ug
Valid period: 90 days
Storage temperature: -20℃
Shipping: at ambient temperature,within a week
Notes: The plasmids shall be transformed into competent cells, and then use after amplification and extraction. Before transformation, please contact technicians or visit the website for the information about the name and concentration of the antibiotics, competent cells and culture temperature.

Handling procedures:
1 dissolving: upon receipt of the freeze dried plasmid, add 20ul of sterile water to the bottom of the ampoule, dissolve the pellets, and sit at room temperature for 1min;
2 mixture (adsorption of plasmids ): mix 200ul competent cells and 5-10ul of plasmid DNA evenly and place it on ice for 30min;

3 heat shock: sit at 42 ℃ for 90s
4 shrink the membrane pore: ice bath for 2min
5 repair cultivation: add 800ul of LB liquid medium to each tube, culture at 37 ℃for 1 hour, 150 r / min;
6 screening cultivation: distribute appropriate (100ul) recovered cells to the LB plate with corresponding antibiotics, place the plate uprightly for 30min (the agar surface must be dried), sit the plate upside down for 12-16h, and colonies appear.
7 Extraction: pick the monoclonal colonies into the LB liquid medium with corresponding antibiotics, shake for 12-16h, and extract the plasmid according to the needs.

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