Intact Genomics (ig^®) chemically competent MG1655 chemically competent cells are suitable for routine DNA transformation and other research purposes. MG1655 is the “wild type” K-12 strain with minimal genetic manipulation having only been cured of the temperate bacteriophage lambda and F plasmid by ultraviolet light and acridine orange (1).

Specifications

Competent cell type:  Chemically competentDerivative of:  MG1655Species:  E. coliFormat:  TubesTransformation efficiency:  ≥5 x 10⁶ cfu/µg pUC19 DNABlue/white screening:  NoShipping condition:  Dry ice

Reagents Needed for One Reaction

ig^® MG1655 chemically competent cells:  50 µlDNA (or pUC19 Control, 10 pg/µl):  1 µlRecovery medium:  1 ml

Storage

ig^® MG1655 competent cells:   -80 ºCpUC19 control DNA:  -20 ºCRecovery medium:  4 ºC

Genotype

F- lambda– ilvG- rfb-50 rph-1

Genomic Features

ig^® MG1655 chemically competent cells have the following genomic features:

• Cured of the temperate bacteriophage lambda and F plasmid
• A frameshift at the end of rph
• ilvG- mutation is a frameshift that knocks out acetohydroxy acid synthase II
• rfb-50 mutation is an IS5 insertion that results in the absence of O-antigen synthesis

Quality Control

Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the high efficiency transformation protocol. Transformation efficiency should be greater than 5×10⁶ CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

Follow these guidelines when using MG1655 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.

Technical SupportIntact Genomics is committed to supporting the worldwide scientific research community by supplying the highest quality reagents. Each new lot of our products is tested to ensure they meet the quality standards and specifications designated for the product. Please follow the instructions carefully and contact us if additional assistance is needed. We appreciate your business and your feedback regarding the performance of our products in your applications.

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