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Team:Macquarie Australia/Protocols/Electroporation
From 2013.igem.org
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<b>(Keep materials on ice!)</b><br><br> | <b>(Keep materials on ice!)</b><br><br> | ||
| - | <b>1)</b>Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube<br><br> | + | <b>1)</b> Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube<br><br> |
| - | <b>2)</b>Add 50µL competent cells (stir very gently to avoid shear)<br><br> | + | <b>2)</b> Add 50µL competent cells (stir very gently to avoid shear)<br><br> |
<img src="https://static.igem.org/mediawiki/2013/4/47/800px-Electroporation_Cuvettes.jpg" alt="Smiley face" align="right" height="300" width="400"> | <img src="https://static.igem.org/mediawiki/2013/4/47/800px-Electroporation_Cuvettes.jpg" alt="Smiley face" align="right" height="300" width="400"> | ||
| - | <b>3)</b>Dry electric cuvette<br><br> | + | <b>3)</b> Dry electric cuvette<br><br> |
<b>4)</b> Pipette 50 µL competent cells to electric cuvette<br><br> | <b>4)</b> Pipette 50 µL competent cells to electric cuvette<br><br> | ||
| - | <b>5)</b>Pulse cuvette<br><br> | + | <b>5)</b> Pulse cuvette<br><br> |
<b><font size = 3>Settings:</font> presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)<br><br></b> | <b><font size = 3>Settings:</font> presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)<br><br></b> | ||
| - | <b>6)</b>Quickly add 500µL LB broth (warmed in hot water bath) to cuvette<br><br> | + | <b>6)</b> Quickly add 500µL LB broth (warmed in hot water bath) to cuvette<br><br> |
| - | <b>7)</b>Mix by gently pipetting up and down<br><br> | + | <b>7)</b> Mix by gently pipetting up and down<br><br> |
| - | <b>8)</b>Working quickly, transfer contents to fresh tube<br><br> | + | <b>8)</b> Working quickly, transfer contents to fresh tube<br><br> |
| - | <b>9)</b>Incubate in warm room under agitation one hour<br><br> | + | <b>9)</b> Incubate in warm room under agitation one hour<br><br> |
Latest revision as of 06:47, 13 September 2013
Electroporation
(Keep materials on ice!)
1) Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube
2) Add 50µL competent cells (stir very gently to avoid shear)
3) Dry electric cuvette
4) Pipette 50 µL competent cells to electric cuvette
5) Pulse cuvette
Settings: presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)
6) Quickly add 500µL LB broth (warmed in hot water bath) to cuvette
7) Mix by gently pipetting up and down
8) Working quickly, transfer contents to fresh tube
9) Incubate in warm room under agitation one hour
