Team:Macquarie Australia/Protocols/SDS Page

From 2013.igem.org

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Pelleted cells were resuspended in 200 uL Milli-Q H2O.
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[[File:800px-Stain_for_SDS-PAGE_gel.jpg|500px|thumb|left|SDS-Page, image by Andra MIhali]]
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<b>1)</b> Resuspend Pelleted cells in 200 uL Milli-Q H2O.
   
   
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Transferred 50 uL of suspension into new Eppendorf tubes and combined with 50 uL 2x TruSep sample buffer.
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<b>2)</b> Transfer 50 uL of suspension into new Eppendorf tubes and combined with 50 uL 2x TruSep sample buffer.
   
   
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Using a Hamilton syringe, the cells were sheared.
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<b>3)</b> Using a Hamilton syringe, shear the cells.
   
   
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Centrifuged the preparations for 3 mins @ 13,000 rpm.
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<b>4)</b> Centrifuge the preparations for 3 mins @ 13,000 rpm.
   
   
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Loaded 20 uL of the supernatant in to the gel.
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<b>5)</b> Load 20 uL of the supernatant in to the gel.
   
   
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Electrophoresis was conducted at constant voltage (200 V) for one hour.
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<b>6)</b> Conduct electrophoresis at a constant voltage (200 V) for one hour.

Latest revision as of 06:29, 13 September 2013


SDS Page

SDS-Page, image by Andra MIhali

1) Resuspend Pelleted cells in 200 uL Milli-Q H2O.


2) Transfer 50 uL of suspension into new Eppendorf tubes and combined with 50 uL 2x TruSep sample buffer.


3) Using a Hamilton syringe, shear the cells.


4) Centrifuge the preparations for 3 mins @ 13,000 rpm.


5) Load 20 uL of the supernatant in to the gel.


6) Conduct electrophoresis at a constant voltage (200 V) for one hour.

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