Intact Genomics (ig®) ccdB Resist™ chemically competent E. coli cells are suitable for high efficiency transformation in a wide variety of applications such as cloning and sub-cloning. ccdB Resist™ cells are capable of propagation of plasmids containing the ccdB gene. Intact Genomics ccdB Resist™ cells provide superb transformation efficiency, allowing for increased opportunity for experimental success.
CUSTOM ALIQUOTS AVAILABLE
Specifications
Competent cell type: Chemically competentSpecies: E. coliFormat: TubesTransformation efficiency: ≥1.0 x 109 cfu/µg pUC19 DNABlue/white screening: YesShipping condition: Dry ice
Reagents Needed for One Reaction
ig® ccdB Resist™ chemically competent cells: 50 µlDNA (or pUC19 Control, 10 pg/µl): 1 µlRecovery medium: 1 ml
Product Includes & Storage
ig® ccdB Resist™ chemically competent cells: -80 ºCpUC19 control DNA: -20 ºCRecovery medium: 4 ºC
Product Benefits
ig® ccdB Resist™ chemically competent cells have the following features:
- Resistance to the ccdB gene product, as well as the T1 and T5 phage
- High transformation efficiency
- endA1 mutation increases plasmid yield significantly
- Capable of blue/white screening
Genotype
F-mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG fhuA::IS2
Quality Control
Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and the high efficiency transformation protocol listed below. Transformation efficiency should be ≥1 x 109 CFU/µg pUC19 DNA.
Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
Follow these guidelines when using ig® ccdB Resist™ chemically competent E. coli.
- 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.
Example Calculation of Transformation EfficiencyTransformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1µg of plasmid into a given volume of competent cells.
TE = Colonies/µg/Dilution
Transform 1 µl of (10 pg/µl) pUC19 control plasmid into 50 µl of cells, add 950 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 100 colonies, the TE is calculated as follows:
Colonies = 100µg of DNA = 0.00001Dilution = 50/1000 x 10/1000 = 0.0005TE = 100/.00001/.0005 = 2.0×10101064-12 1064-24
