---+ Checking Transformation Efficiency of Chemically Competent Cells Adapted from _Molecular Cloning: A Laboratory Manual 3rd Ed._, Sambrook and Russell (2001) ---++ SUPPLIES: ---+++ Equipment: * Shaking Incubator or Shaking Platform and 1L flask clamps * 42°C Heating Bath or Block * 37°C Incubator * Colony Counter ---+++ Consumables: * Agar plates with appropriate antibiotic ---+++ Buffers and Solutions: * Competent cells prepared using the [[ProtocolsChemCompCellsInoue][Inoue method]] * 10pg / μL solution of a standard plasmid (e.g. pUC19) * [[ProtocolsRecipesSOB][SOC Medium]] ---++ PROTOCOL: 1 ) Transform 10pg of a standard plasmid (e.g. pUC19) in 50μL of cells using the standard <span style="color: #0000ff; text-decoration: underline">[[ProtocolsTransformingChemCompCells][transformation protocol]]</span> 1 ) Plate 50uL of transformed cells in triplicate on appropriate antibiotic 1 ) Grow plates overnight, and count colonies the next morning 1 ) Average colony counts for the three plates; efficiency = (# colonies) x 10<sup>6</sup> cfu / μg 1 ) Expected efficiency for the Inoue protocol is 1-3 x 10<sup>8</sup> colonies / μg of plasmid -- Main.KateElston - 25 Oct 2018
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Topic revision: r1 - 2018-10-25 - KateElston