Team:UCL/Project/Protocols

From 2013.igem.org

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<p class="minor_title">Stable Transfection Of Adherent Cells</p>
<p class="minor_title">Stable Transfection Of Adherent Cells</p>
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This protocol is for the stable transfection of eukaryotic adherent cell types in a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible.
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This protocol is for the stable transfection of eukaryotic adherent cell types in a single well of a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible. When transfecting multiple wells, make a 'master mix' with 10% of
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<b>2)</b> Incubate cells in their normal growth conditions (37^0 C and 5% CO2).
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<b>2)</b> Incubate cells in their normal growth conditions (37^0 C and 5% CO2)for 24 hours.
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<b>3)</b>  
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<b>3)</b> Dilute 2µg of DNA dissolved in TE buffer (min conc. 0.1µg/µl) with serum, protein and antibiotic free medium (to avoid macromolecular interference with complex formation) to a total of 100µl. Mix.
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<b>4)</b> Add 10µl
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Revision as of 11:45, 25 September 2013

There are two solutions to \(ax^2 + bx + c = 0\) and they are $$x = {-b \pm \sqrt{b^2-4ac} \over 2a}.$$

Bacterial Lab Protocols

Stable Transfection Of Adherent Cells

This protocol is for the stable transfection of eukaryotic adherent cell types in a single well of a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible. When transfecting multiple wells, make a 'master mix' with 10% of

1) The day before transfection, seed 0.9-4x10^5 cell per well of the six well plate with 2ml of appropriate growth medium. This should produce a confluence of 40-80% for the next day's transfection.

2) Incubate cells in their normal growth conditions (37^0 C and 5% CO2)for 24 hours.

3) Dilute 2µg of DNA dissolved in TE buffer (min conc. 0.1µg/µl) with serum, protein and antibiotic free medium (to avoid macromolecular interference with complex formation) to a total of 100µl. Mix.

4) Add 10µl

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Mammalian Lab Protocols

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