Difference between revisions of "Team:Paris Bettencourt/Protocols/Gel Electrophoresis"

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       <li>Preparation of the Gel (50mL)
 
       <li>Preparation of the Gel (50mL)
 
         <ul>
 
         <ul>
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           <li>Weight 50mg of Agarose NEEO.</li>
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           <li>Mix 0.5 grams of agarose with 50mL 0.5% TAE solution</li>
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          <li>put the 50mg of Agarose in an Erlenmeyer and add 50mL of TAE 0.5%.</li>
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           <li>Heat the Erlenmeyer in a microwave until the liquid becomes totally transparent.</li>
 
           <li>Heat the Erlenmeyer in a microwave until the liquid becomes totally transparent.</li>
 
           <li>Let it rest until you can take the Erlenmeyer in your hand without pain for 10 seconds.</li>
 
           <li>Let it rest until you can take the Erlenmeyer in your hand without pain for 10 seconds.</li>
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           <li>Add 5uL of SYBR® Safe and mix gently until the liquid takes and uniform reddish color.</li>
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           <li>Add 5uL of SYBR® Safe (10000X) and mix gently until the liquid takes and uniform reddish color.</li>
 
           <li>Pour the liquid in a gel mold with the desired number of well and wait 20-30 min (or until the gel is solidified).</li>
 
           <li>Pour the liquid in a gel mold with the desired number of well and wait 20-30 min (or until the gel is solidified).</li>
 
           <li>Put the gel inside the electrophoresis machine and pour some TAE 0.5% until the gel is totally submerged.</li>
 
           <li>Put the gel inside the electrophoresis machine and pour some TAE 0.5% until the gel is totally submerged.</li>

Revision as of 16:38, 6 September 2015

Gel-Electrophoresis protocol
  • Preparation of the Gel (50mL)
    • Mix 0.5 grams of agarose with 50mL 0.5% TAE solution
    • Heat the Erlenmeyer in a microwave until the liquid becomes totally transparent.
    • Let it rest until you can take the Erlenmeyer in your hand without pain for 10 seconds.
    • Add 5uL of SYBR® Safe (10000X) and mix gently until the liquid takes and uniform reddish color.
    • Pour the liquid in a gel mold with the desired number of well and wait 20-30 min (or until the gel is solidified).
    • Put the gel inside the electrophoresis machine and pour some TAE 0.5% until the gel is totally submerged.
  • Preparation of the samples
    • Mix 5uL of each sample with 1uL of 6X loading dye in microcentrifuge tubes/ on a piece of parafilm / in PCR tubes.
  • Pipette 5-6uL of each sample inside a different well.
  • Pipette 5uL of the appropriate ladder in one of the wells.
  • Close the machine and launch the migration with the desired voltage
  • After the desired migration time turn of the machine, remove the lid and put the gel inside the Blue light revelator.