GoldBio/JM109 Chemically Competent E. coli Cells/10 x 50 μl/CC-114-10x50

GoldBio’s JM109 Chemically Competent E. coli cells are suitable for high efficiency transformation in a wide variety of routine applications such as plasmid isolation, library construction, cloning, and subcloning. JM109 is a K strain bacterium that carries multiple features including the ?80lacZ?M15 marker, which provides α-complementation of the β-galactosidase gene with blue/white screening protocol as well as the hsdR17 genotype which prevents endogenous endonucleases from cleaving DNA.

Product SpecificationsCompetent cell type: Chemically competentDerivative of: JM109Species: E. coliFormat: TubesTransformation efficiency: ≥6 x 10⁵ cfu/μg pUC19 DNABlue/white screening: Yes

Storage/Handling: This product may be shipped on dry ice. JM109 Chemically Competent E. colicells should be stored at -80°C, pUC19 Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.

endA1, recA1, gyrA96, thi, hsdR17 (rk^–, mk⁺), relA1, supE44, Δ( lac-proAB), [F′ traD36, proAB, lacI^qZΔM15]

Reagents Needed for One Reaction

• JM109 chemically competent cells: 50 μl
• DNA (or pUC19 Control, 10 pg/μl): 1 μl
• Recovery medium: 1 ml

Quality ControlTransformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥6 x 10⁵ CFU/μg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

• Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
• Thaw competent cells on ice, and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.

Calculation of Transformation EfficiencyTransformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1 μg of plasmid into a given volume of competent cells.

• TE = Colonies/μg/Dilution
  • Colonies = the number of colonies counted
  • μg = amount of DNA transformed in μg
  • Dilution = total dilution of the DNA before plating

Example: Transform 1 μl of (10 pg/μl) control plasmid into 25 μl of cells, add 975 μl of Recovery Medium. Dilute 10 μl of this in 990 μl of Recovery Medium and plate 50 μl. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:Colonies = 250μg of DNA = 0.00001 Dilution = 10/1000 x 50/1000 = 0.0005 TE = 250/0.00001/0.0005 = 5.0 × 10¹⁰