- Description Intact Genomics (IG®) Recombineering Electrocompetent Cells are designed for high-efficiency transformation in a wide range of applications, including DNA, plasmid, and BAC engineering, as well as homologous recombination. This strain is a SW102 derivative containing a defective lambda prophage in a DH10B [λcI857 ind1 (cro-bioA<>tet)] background. In addition, it carries a fully functional gal operon except for a galK deletion, which enables efficient DNA or BAC modification through galK positive/negative selection. This strain is tetracycline resistant (5 µg/mL). Specifications: Competent cell type: Electrocompetent Derivative of: SW102 Species: E. coli Format: Tubes Transformation efficiency: ≥ 1.0 x 1010 cfu/µg pUC19 DNA Shipping condition: Dry ice Reagents Needed for One Reaction: IG® Recombineering Electrocompetent Cells: 50 µl DNA (or pUC19 Control, 10 pg/µl): 1 µl Recovery Medium: 1 ml Product Components and Recommended Storage Condition: IG® Recombineering Electrocompetent Cells: -80 ºC pUC19 control DNA: -20 ºC Recovery Medium: 4 ºC Genomic Features and Benefits: IG Recombineering Electrocompetent Cells have the following features: This strain carries an engineered, heat-inducible prophage in which red genes (exo, bet, gam) are placed under the control of the heat-inducible promoter pL, which is repressed at 32 °C and induced at 42 °C. It is a ΔgalK derivative. It supports galK positive/negative selection for precise, scarless engineering. TetR: The cro-bioA region has been replaced with a tetracycline resistance gene (tetra, 5 µg/ml). Genotype: DH10B [λcI857ind1 (cro-bioA<>tet)] 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 1010 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity. General Guidelines: Follow these guidelines when using IG Recombineering 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. 1266 1266-12 1266-48
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