EPI400 electro competent cells (MOFY-0822-FY474)


Cat: MOFY-0822-FY474
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Specifications

DescriptionEPI400 electroporation of competitive cells can only be used for electroshock transformation, not for heat shock transformation. The strain is derived from the EC100 strain. After deleting the pcnB gene that controls the plasmid copy number in the EC100 nuclear gene, an inducible promoter-driven pcnB gene is introduced, which is the EPI400 strain. The EPI400 strain can reduce the copy number of plasmids, especially suitable for the cloning of various unstable DNA or virulence genes. After adding WDEPI-inducer I, the plasmid yield can be increased to a normal state.
Gene TypesF- mcrA Δ (mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ (ara, leu)7697 galU galK λ- rpsL (StrR) nupG trfA tonA pcnB4 dhfr
Size50 µL/vial
QC resultTransformation efficiency of pUC19 plasmid detection:> 1×1010 cfu/μg DNA
Note1. When adding DNA, the volume should not be greater than 1/10 of the competent volume.
2. When adding competitive cells into the electric shock cup, avoid generating air bubbles, which will increase the risk of arc discharge.
3. When the DNA is impure or contaminated with salt, ethanol, protein and buffer, the transformation efficiency drops sharply.
4. The ions in the shock cup can increase the conductance of the solution, increasing the risk of electric current and arcing in solutions containing cells and DNA.
5. If you want to transform large plasmids or want to obtain higher transformation efficiency, it is recommended to use a high-purity plasmid extraction kit to extract the plasmids. Double the size of the plasmid, and the transformation efficiency drops by an order of magnitude.
6. For the transformation of ligation products, try to ethanol-precipitate the DNA before transformation and resuspend the product with an appropriate amount of TE buffer (10 mM Tris HCl, pH7.5; 1 mM EDTA) to ensure that the DNA concentration does not exceed 100 ng/μl. Too high a concentration of ligation product or too large a volume of ligation product will reduce conversion efficiency and increase the risk of arcing.
7. Be gentle when mixing plasmids. Do not use a pipette tip that is too small when absorbing compatible cells (ordinary 200ul pipette tips should be cut off by 0.6cm). Avoid excessive force, so as not to damage the cell membrane due to excessive shearing force and reduce the Conversion efficiency. Transforming high concentrations of plasmids or ligation products can correspondingly reduce the amount of bacteria that are ultimately used for plating.
8. Electroporation of competitive cells should be kept below -80°C as much as possible. Over-storage at temperatures above -80°C will cause the transformation efficiency to decrease.

Properties

ShippingDry ice
StorageStore at -80 °C
Handling AdviceImmediately upon receipt, cells were transferred from dry ice to -80°C refrigerator.
Related Products
MOFY-0822-FY475EPI400 chemically competent cells (MOFY-0822-FY475)
For Research Use Only | Not For Clinical Use.
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