Team:Macquarie Australia/Protocols/Electroporation

From 2013.igem.org

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<b>5)</b>Pulse cuvette<br><br>
<b>5)</b>Pulse cuvette<br><br>
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<b><font size = 3>Settings:</font></b> presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)<br><br>
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<b><font size = 3>Settings:</font> presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)<br><br></b>
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Quickly add 500µL LB broth (warmed in hot water bath) to cuvette<br><br>
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Mix by gently pipetting up and down<br><br>
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<b>6)</b>Quickly add 500µL LB broth (warmed in hot water bath) to cuvette<br><br>
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Working quickly, transfer contents to fresh tube<br><br>
+
 
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Incubate in warm room under agitation one hour<br><br>
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<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>9)</b>Incubate in warm room under agitation one hour<br><br>

Revision as of 06:46, 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)

Smiley face 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

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