Team:CSU Fort Collins/Notebook/Protocols=Gel

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           <li>Using a balance, mass out 1.0 gram of agarose. Mix this with 100 MICROL of TAE 1X Buffer.</li>
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           <li>Using a balance, mass out 1.0 gram of agarose. Mix this with 100 mL of TAE 1X Buffer.</li>
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           <li>Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.<li>
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           <li>Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.</li>
           <li>Pour the mixture into a gel plate and insert an 8-lane comb. Allow to cool and solidify in the fridge or on the benchtop</li>
           <li>Pour the mixture into a gel plate and insert an 8-lane comb. Allow to cool and solidify in the fridge or on the benchtop</li>
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           <li>Mix your samples to run in the gel. Use 10 MICROL of the DNA Ladder combined with 2 MICROL SYBR Green in the first lane and for all samples, mix 5 MICROL of sample with 5 MICROL of nuclease-free water and 2 MICROL of SYBR Green/Dye Mixture.</li>
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           <li>Mix your samples to run in the gel. Use 10 &#956;L of the DNA Ladder combined with 2 &#956;L SYBR Green in the first lane and for all samples, mix 5 &#956;L of sample with 5 &#956;L of nuclease-free water and 2 &#956;L of SYBR Green/Dye Mixture.</li>
           <li>Place gel in tray into the gel electrophoresis apparatus; Wells should be located towards the negative (conventionally black) end. Fill apparatus with 1X TAE until the gel is completely submerged. Do not overfill.</li>
           <li>Place gel in tray into the gel electrophoresis apparatus; Wells should be located towards the negative (conventionally black) end. Fill apparatus with 1X TAE until the gel is completely submerged. Do not overfill.</li>
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           <li>Load 10 MICROL of each sample into the appropriate wells.</li>
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           <li>Load 10 &#956;L of each sample into the appropriate wells.</li>
           <li>Connect wiring and run gel electrophoresis for 1 hour.</li>
           <li>Connect wiring and run gel electrophoresis for 1 hour.</li>
           <li>Analyze gel in UV light and see if samples match the expected size using the ladder as a guide.</li>
           <li>Analyze gel in UV light and see if samples match the expected size using the ladder as a guide.</li>
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Latest revision as of 17:50, 17 October 2014

CSU iGEM 2014

Gel Electrophoresis Protocol

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Gibson Assembly
Cloning Genes
Plasmid Miniprep
Yeast DNA Extraction
Gel Electrophoresis
PCR Product Purification

  1. Using a balance, mass out 1.0 gram of agarose. Mix this with 100 mL of TAE 1X Buffer.
  2. Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.
  3. Pour the mixture into a gel plate and insert an 8-lane comb. Allow to cool and solidify in the fridge or on the benchtop
  4. Mix your samples to run in the gel. Use 10 μL of the DNA Ladder combined with 2 μL SYBR Green in the first lane and for all samples, mix 5 μL of sample with 5 μL of nuclease-free water and 2 μL of SYBR Green/Dye Mixture.
  5. Place gel in tray into the gel electrophoresis apparatus; Wells should be located towards the negative (conventionally black) end. Fill apparatus with 1X TAE until the gel is completely submerged. Do not overfill.
  6. Load 10 μL of each sample into the appropriate wells.
  7. Connect wiring and run gel electrophoresis for 1 hour.
  8. Analyze gel in UV light and see if samples match the expected size using the ladder as a guide.



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