pUC19|353861 |BNCC

Essential Information Certificate Related Products

pUC19 plasmid

Subculture procedure	①Dissolution: Add 20 μL sterile water to lyophilized plasmid powder and dissolve, let stand at Room temperature for 1 min; ② Mix: Combine 50 μl competent cells with 10 μl plasmid DNA, mix thoroughly, and incubate on ice for 30 min; ③ Heat shock transformation: Incubate at 42°C for 90 s; ④ Shrink membrane pores: Ice bath for 2 min; ⑤ Repair culture: Add 800 μl liquid medium to each tube, incubate on a 37°C shaking incubator for 1 h (150 rpm); ⑥ Screening culture: Spread an appropriate volume (100 μl) of the revived cells onto LB plates with the corresponding antibiotic. Incubate plates upright for 30 min (ensure agar surface is dry), then invert and incubate for 12-16 hours until colonies appear; ⑦ Extraction: Pick a single colony into the corresponding antibiotic-containing LB liquid medium. Incubate with shaking for 12–16 hours. Extract the plasmid as required for the experiment.
Growth conditions	LB + antibiotic, 37°C
Storage conditions	2-8°C
Safety level	1
Sharing mode	Public welfare sharing

Product format: freeze dried, 1.0-2.0ug
Validity period: 90 days
Storage temperature: -20℃
Shipping way: at ambient temperature,within one 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.

Download certificate