DH5alpha 感受态细胞(化转)/ DH5alpha

DH5α chemically Competent Cells

Packing 

DH5α锛?nbsp;                                                     100ul/tube
pUC19 (control vector锛?0pg/ul):                   10ul
Storage condition锛?nbsp;                           -80鈩冿紙6month锛?nbsp;

DH5α chemically Competent Cells Genotype
F- φ80 lac ZΔM15 Δ(lacZYA-arg F) U169 endA1 recA1 hsdR17(rk-,mk+) supE44λ- thi -1 gyrA96 relA1 phoA

DH5α chemically Competent Cells Description

DH5 alpha strain is the most commonly used competent cell in laboratory. The deletion of endonuclease (endA) increased the yield and quality of plasmid DNA, and the recombinant enzyme deficiency type (recA) reduced the homologous recombination probability of the inserted fragments to ensure the stability of the insertion of DNA, and the existence of lacZ Delta M15 could be used for the selection of blue and white leukoplakia by DH5 alpha. DH5 pUC19 competent cells were produced by special technology, and the efficiency >5×108 cfu/μg DNA.

DH5α chemically Competent Cells transformation

DH5α chemically Competent Cells transformation

1. Please store DH5α chemically Competent Cells in -80°C for late use. Place it on Room temperature to partial melting.Then Thaw a tube of DH5α chemically Competent Cells锛?00μl 锛塷n ice for 25min.

2. Add 1µl plasmid DNA (not more than 10ul) to the cell mixture. Carefully flick the tube 4–5 times to mix cells and DNA. Do not vortex.Place the mixture on ice for 25 minutes,Do not mix. Place it in the 42°C water,then immediately transfer the tube back into ice for 2min. Do not mix.

3. Add 700ul LB or YEB liquid nutrient medium. then put on the Constant temperature shaker (200 rpm) for 1 hours.

4. Pellet the mixture by centrifugation at 5000rpm/min for 1min. Pour off about 100ul of cell supernatant, then resuspend the cell pellet in the remaining medium by gently vortexing the tube. Spread the cell on the LB or YEB Resistance plates by antibiotic, then inverted and Incubate at 37°C. overnight.