PRODUCT INFORMATION

DynaCompetent Cells

Product Name :JM109 for Electroporation(Previous name: Electrocompetent Cell JM109)
Code No. :DS218

Size :

50 μl × 5

Competency :> 2 × 109 cfu/μg

Supplied product :SOC medium, 1 ml × 5(50 μl cell, pUC19)
This product is for research use only

Description :DynaCompetent Cells

JM109 for Electroporation is designed for high efficiency transformation byelectroporation and is suitable for DNA library construction and all kinds of cloning procedures. E. coli
strain JM109 is one of the standard competent cells for molecular biology applications. The JM109 cellcontains laqIqZΔM15 gene on F’ episome, which allows blue-white color screening of transformants.
Genotype of E. coli strain JM109 : recA1, supE44, endA1, hsdR17, gyrA96, relA1, thi, Δ(lac-proAB),F′ [traD36, proAB+, laqIqZΔM15]
Quality control :Transformation by electroporation was carried out according to the method described in this Product
Information using supercoiled pUC19 plasmid. Transformants were plated on LB plates containing 50
μg/ml ampicillin. The efficiency was confirmed to be greater than 2 × 109 cfu/μg with 50 μl of theelectrocompetent cells.
Storage condition :Stable at -80℃ with little or no loss in transformation efficiency for 12 months from the date of receipt.
Upon receipt, store the DynaCompetent Cells JM109 for Electroporation in a freezer at -80oC directly from a dry
ice shipping box and store SOC medium at room temperature or at -80oC.
Handling of competent cells :• After thawing electrocompetent cells on ice, cells tend to lose transformation efficiency gradually.
Transformation should be started immediately following thawing cells on ice.
• Electroporation is temperature-dependant. Electrocompetent cells should be maintained at 0-4°C before
pulsing with electroporation apparatus.
• Use of refrozen electrocompetent cells is not recommended.
Composition of SOC medium supplied :20 g/L5 g/L
0.5 g/L
0.186 g/L
2.4 g/L
4 g/L

tryptoneyeast extract
NaCl
KCl
MgSO4, anhydrous
glucose1
Ver. 3.0

PRODUCT INFORMATION

Transformation (electroporation) procedure :● Materials to be supplied by user
• Electroporation apparatus• Sterile electroporation cuvettes (0.1 cm gap)

• LB plates with antibiotic• Sterilized pasteur pipettes• Sterile 14 ml polypropylene culture tubes• 37°C shaker• Sterile spreaders• 37°C incubator

If blue-white screening is required to select transformants,• 20 mg/ml X-Gal in dimethylformamide (DMF)• 100 mM IPTG in water (filter sterilized)● Transformation
Before start:• Chill sterile electroporation cuvettes (0.1 cm gap) on ice.
* Electrocompetent cells should be maintained at 0-4°C before pulsing.
• Place 14 ml sterile polypropylene culture tubes containing 950 μl of SOC medium (at roomtemperature) for each electroporation near the electroporation apparatus.
* It is important to add SOC medium to electrocompetent cells immediately after pulsing.
• Set up the electroporation apparatus according to the manufacturer’s instructions.
1. Thaw the electrocompetent cells on ice and add DNA solution*(1-3 μl) to the cells then flick gently.
* DNA solution*: Highly purified DNA is recommended. DNA should be dissolved in pure water or
1×TE. For ligation mixture, see “To use ligation mixture for electroporation” below.
2. Quickly transfer the mixture of cells and DNA to a chilled electroporation cuvette, and tap the mixture
to the bottom of the cuvette.
* Electroporation should be performed immediately after mixing electrocompetent cells and DNA.
3. Place the cuvette into the electroporation apparatus, then pulse.
4. Remove the cuvette from the electroporation apparatus and immediately add the 950 μl of SOC
medium to the cuvette with a sterile pasteur pipette to suspend cells.
5. Transfer the cell suspension to the sterile 14 ml polypropylene culture tube. Culture the cell at 37°C
for 1 hr in a shaker.
6. Spread aliquot of the cell to an LB agar plate containing appropriate antibiotic.
If blue-white color screening is required, spread 25 μl of 20 mg/ml X-Gal and 50 μl of 100 mM
IPTG on the LB agar plates and allow these reagents to absorb 30 minutes prior to inoculating cells.
7. Incubate the plate at 37°C overnight.

2Ver. 3.0

PRODUCT INFORMATION● To use ligation mixture for electroporation
DNA for electroporation should be highly purified and dissolved in pure water or low-salt buffer (e.g.
1 × TE). Ligation mixture contains salts and impurities that interfere with electroporation. Highconcentration of salts in the mixture of cells and DNA often causes arcing during electroporation. To use
ligation mixture for electroporation, DNA should be purified by phenol/chloroform extraction following
ethanol precipitation or with a spin column. The phenol/chloroform extraction following ethanol
precipitation gives successive results for ligation mixture prepared with DNA Ligation Kit ver. 2 (DS110).

Reference:
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, NY.

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3Ver. 3.0