pGEX-4T-1|394099 |BNCC

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pGEX-4T-1

Growth conditions	37℃； 18-24h; aerobic
Subculture procedure	① Dissolve: Add 20 μ L of sterile water to the plasmid powder and let it stand at Room temperature for 1 minute; ② Mixing: Mix 200 μ l of competent cells with 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 of liquid culture medium to each tube and culture on a shaker at 37 ℃ for 1 hour (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 a petri dish 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.
Storage conditions	2-8 ℃
Sharing mode	Public welfare sharing