Intact Genomics (ig^®) ER2738 Phage Display ElectroCompetent Cells are suitable for protein expression and preparation of antibody or peptide phage display libraries.  ER2738 cells are also useful for protein expression, M13 phage work, general cloning, and blue/ white screening.

Custom Aliquots Available

Specifications

Competent cell type:  ElectroCompetentSpecies:  E. coliFormat: TubesTransformation efficiency:  ≥ 4 x 10¹⁰ cfu/µg pUC19 DNABlue/white screening:  YesShipping condition:  Dry ice

Reagents Needed for One Reaction

ig^®  ER2738 Phage Display electrocompetent cells:  25 µlDNA (or pUC19 Control, 10 pg/µl):  1 µlRecovery medium:  1 ml

Storage

ig^® ER2738 Phage Display electrocompetent cells:  -80 ºCpUC19 control DNA:  -20 ºCRecovery medium:  4 ºC

Genomic Featuresig^® ER2738 phage display electrocompetent cells have the following features:

• >4 x 10¹⁰cfu/µg efficiency with electroporation.
• Amber suppressor strain (glnV)

Genotype

[F’proA+B+ lacIq Δ(lacZ)M15 zzf::Tn10 (tetr)] fhuA2 glnVΔ(lac-proAB) thi-1Δ(hsdS-mcrB)5

Quality Control

Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥5 x 10¹⁰ CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity. 

General Guidelines

Follow these guidelines when using ig^® ER2738 Phage Display electrocompetent cells:

• 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.

Note: A high-voltage electroporation apparatus such as Bio-Rad Gene Pulser II #165-2105, capable of generating field strengths of 16 kV/cm is required.

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 200 colonies, the TE is calculated as follows:

Colonies = 200µg of DNA = 0.00001Dilution = 50/1000 x 10/1000 = 0.0005TE = 200/.00001/.0005 = 4.0×10¹⁰1217-12 1217-24