EPI400 感受态细胞(化转)/ EPI400 chemi

EPI400 chemically Competent Cells

 

EPI400 Chemically Competent Cell Components

EPI400锛?nbsp;                                   100μl/tube

pUC19 (control vector锛?0pg/μl):                    10μl

Storage at                             -80鈩冿紙6month锛?/p>

 

EPI400 Chemically Competent Cell Genotype

F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG trfA tonA pcnB4 dhfr

 

EPI400 Chemically Competent Cell Description

EPI400 is derived from EC100 strain. After deletion of pcnB gene that controls plasmid copy number in EC100 nuclear gene, it is introduced into a pcnB gene driven by promoter, EPI100 strain. EPI400 strain can reduce the copy number of plasmid, especially suitable for all kinds of unstable DNA or virulence gene cloning. After inducer CopyCutter Induction Solution, it can increase plasmid yield to normal state. [mcrA, Delta (mrr-hsdRMS-mcrBC)] genotypes make EPI400 strains suitable for cloning DNA rich in methylcytosine or methyl adenine. The mutation of recA1 and endA1 is beneficial to the insertion of the stable and high purity plasmid DNA of DNA. The presence of lacZ Delta M15 markers can enable EPI400 to be used in the screening of blue leukoplakia, and tonA gives its ability to resist phage T1 and T5, and rpsL endows streptomycin to its resistance. The EPI400 receptive cells were made by special technology, and the conversion efficiency of pUC19 plasmid detection could reach 5 * 107 cfu/ mu g DNA.

 

EPI400 Chemically Competent Cell Transformation Protocol
1. Please store EPI400 Chemically Competent Cell in -80°C for late use.
Take it then insert the tube in the ice immediately for 5 minutes.
2. Add your plasmid DNA (vector) 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.
3. Place it in the 42°C water for 45 seconds,then immediately transfer the tube back into ice for 2min. Do not mix.
4. Add 700ul LB or 2YT liquid nutrient medium (No antibiotic). then put on the Constant temperature shaker (200 rpm) for 1 hours at 37°C.
5.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 2YT Resistance plates by antibiotic, then inverted and Incubate overnight at 37°C.