(r1) Barrick Lab :: ProtocolsTestingTransformEff

---+ 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 / &mu;L solution of a standard plasmid (e.g. pUC19)
   * [[ProtocolsRecipesSOB][SOC Medium]]

---++ PROTOCOL:

   1 ) Transform 10pg of a standard plasmid (e.g. pUC19) in 50&mu;L 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 / &mu;g
   1 ) Expected efficiency for the Inoue protocol is 1-3 x 10<sup>8</sup> colonies / &mu;g of plasmid




-- Main.KateElston - 25 Oct 2018

Barrick Lab > ProtocolList > ProtocolsTestingTransformEff

Contributors to this topic

KateElston, JeffreyBarrick

Topic revision: r1 - 2018-10-25 - 21:52:44 - Main.KateElston