Intact Genomics (ig^®) chemically competent BL21(DE3)pLysS chemically competent cells are suitable for transformation and routine protein expression. The pLysS plasmid produces T7 lysozyme that reduces base-level expression of the gene of interest, subsequently allowing for tighter control of expression and is, therefore, suitable for the expression of toxic genes.  BL21(DE3)pLysS chemically competent cells also carry the chloramphenicol resistance gene.

Specifications

Competent cell type:  Chemically competentDerivative of:  BL21(DE3)pLysSSpecies:  E. coliFormat:  TubesTransformation efficiency:  ≥3 x 10⁷ cfu/µg pUC19 DNABlue/white screening:  YesShipping condition:  Dry ice

Reagents Needed for One Reaction

ig^® BL21(DE3)pLysS chemically competent cells:  50 µlDNA (or pUC19 Control, 10 pg/µl):  1 µlRecovery medium:  1 ml

Storage

ig^® BL21(DE3)pLysS competent cells:   -80 ºCpUC19 control DNA:  -20 ºCRecovery medium:  4 ºC

Genotype

F–pLysS, Cm^r ompT hsdS(rB– mB–) gal dcm λ(DE3)

Genomic Features

ig^® BL21(DE3)pLysS chemically competent cells have the following features:

• T7 Expression Strain
• Deficient in both lon (1) and ompT proteases
• Resistant to phage T1 (fhuA2)
• B Strain
• Suitable for expression of toxic genes

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 3×10⁷ CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

Follow these guidelines when using BL21(DE3) pLysS 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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