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CompetentPro™ Kit (EC1001)

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Quick Overview

CompetentProTM Kit (EC1001) allows users to produce highly efficient chemically competent cells whose titer will match original commercial E. Coli user bought from the companies. With CompetentProTM Kit, any chemical competent cell user has used can be applied to get very effective chemically competent E. Coli which quality controlled for a transformation efficiency of 1 x 109 cfu/µg plasmid DNA. The prepared E. Coli by this kit are ideal for high-efficiency cloning and plasmid propagation which maximizes cloning efficiency in single-tube format and enhances genomic DNA cloning capabilities. Scientists in ATCGbio Life Technology Inc. have used this kit to prepare E. Coli for highly demanding applications including lentiviral backbone plasmids construction and shRNA viral plasmids construction. With this product, we no longer have to purchase commercial expensive chemically competent E. Coli, and we are sure that you don’t have to either. CompetentProTM Kit provides enough reagents to make 5 x 40 tubes for single-tube transformation format, total 200 tubes in cost of $1 for each tube. It just takes about 4-7 hours for the preparation.

Kit Components

LB supplement  (EC1001-01)          2.0ml in 2ml tube x 1

Storage solution  (EC1001-02)        2.0ml in 2ml tube x 5

CompetentPro™ Kit (EC1001)


Following the procedure is for preparing 40 single-tube formats E. Coli. The kit contains enough reagents performing 5 times of this procedure.

Required Materials and Equipments

  • Chemically competent E. Coli provided by users
  • 40ml LB medium for 40 tubes preparation
  • 50ml sterile centrifuge tubes
  • 4°C centrifuge capable to hold 50.0ml tubes and 1500 x g force
  • 4°C cold room
  • 40 sterile microtubes (1.7ml or 2.0ml)
  • Spectrophotometer capable to measure absorbance at 600nm with 1cm light path
  • E. Coli incubator capable to rotate at 220rpm, 37 °C


  1. Culture your E. Coli at 2 ml LB medium overnight
  2. Add 40 ml LB medium into 50 ml sterile tube
  3. Add 0.5 ml overnight cultured E. Coli
  4. Add 0.4 ml provided LB supplement solution
  5. Incubate at 37 °C with 220 rpm for 4-7 hours until absorbance at 600 nm (OD 600) reaches at 0.3-0.5

To get highest transformation efficiency, following steps should be performed in cold room (4°C).

  1. Prepare the storage box and ice buckets with ice in cold room.
  2. Insert the tube containing incubated 40ml E. Coli into ice for 10 min.
  3. Centrifuge the tube at 1500 x g  for 10min at 4°C.
  4. Carefully decant supernatant and leave the tube upside-down for 2-3min.
  5.  Resuspend the E. Coli pellet with 1ml storage solution and disperse well by using a sterile glass rod or heat-sterilized Pasteur pipet (Do not vortex or pipet up-down).
  6.  Add another 1ml of storage solution. Mix well with a sterile rod. Keep on ice for another 20min.
  7.  Aliquot out 50µl E. Coli solution each into microtubes.
  8.  Transfer into the storage box and store at -80°C.


The capabilities and characteristics of competent cells prepared by CompetentProTM Kit (EC1001) will be very similar to the ones originally purchased from companies. For this reason, the transformation procedures should be same as what described in the original instruction of chemically competent E. Coli. Typically, the procedures will be 10-20 min incubation with plasmid on ice followed by 1min heat-shock at 42°C and 2 min incubation on ice. After addition of 0.5 ml SOC medium, the cells will be incubated for 1 hr at 37 °C in 220 rpm., then all or aliquot will be separated on an appropriate antibiotics plate.

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