Difference between revisions of "Ellis:Back Door/Protocols/Competent Cells"

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(Replacing page with '=Making Electrocompetent E. coli Cells=')
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=Making Electrocompetent E. coli Cells=
=Making Electrocompetent E. coli Cells=
<li>Get required amount of electrocompetent cells from the -80°C freezer and let them thaw slowly on ice. Usually 1 tube (50μL / tube) per transformation.</li>
<li>Take the appropriate amount of [http://www.bio-rad.com/webroot/web/images/lsr/products/gene_transfer_rnai/product_detail/global/lsr_gp_cuvettes.jpg electroporation cuvettes] from the -20°C freezer (1 per transformation) and label them accordingly</li>
<li>Place 1-2μL of the DNA you wish to transform against the side of the cuvette, between the electrodes.</li>
<li>Add your competent cells to each transformation by targeting the drop of DNA previously deposited.</li>
<li>Keeping the cuvettes on ice, add 50μL of the competent cells and mix by pipetting up and down trying to avoid creating air bubbles</li>

Revision as of 02:26, 14 June 2011

Making Electrocompetent E. coli Cells