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Team:CSU Fort Collins/Notebook/Protocols=Gel
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<li><a href='/Team:CSU_Fort_Collins/Members/'><span>Members</span></a></li> | <li><a href='/Team:CSU_Fort_Collins/Members/'><span>Members</span></a></li> | ||
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<li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/DailyNotes'><span>Daily Notes</span></a> | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/DailyNotes'><span>Daily Notes</span></a> | ||
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| - | <li><a href=" | + | <li><a href="/Team:CSU_Fort_Collins/Notebook/Biosensor/Jun"><span>June</span></a></li> |
| - | <li><a href=" | + | <li><a href="/Team:CSU_Fort_Collins/Notebook/Biosensor/Jul"><span>July</span></a></li> |
| - | + | <li class='last'><a href='/Team:CSU_Fort_Collins/Notebook/Biosensor/Aug'><span>August</span></a></li> | |
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<ul> | <ul> | ||
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| - | <li class='last'><a href=" | + | <li class='last'><a href="/Team:CSU_Fort_Collins/Notebook/Breakdown/Sep"><span>September</span></a></li> |
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<li><a href='/Team:CSU_Fort_Collins/Collab/'><span>Collaboration</span></a></li> | <li><a href='/Team:CSU_Fort_Collins/Collab/'><span>Collaboration</span></a></li> | ||
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| - | <li>Using a balance, mass out 1.0 gram of agarose. Mix this with 100 | + | <li>Using a balance, mass out 1.0 gram of agarose. Mix this with 100 mL of TAE 1X Buffer.</li> |
<li>Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.</li> | <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> | ||
| - | <li>Mix your samples to run in the gel. Use 10 | + | <li>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.</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> | ||
| - | <li>Load 10 | + | <li>Load 10 μL 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> | ||
Latest revision as of 17:50, 17 October 2014
Gel Electrophoresis Protocol
Show Table of Contents
- Using a balance, mass out 1.0 gram of agarose. Mix this with 100 mL of TAE 1X Buffer.
- Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.
- 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
- 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.
- 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.
- Load 10 μL of each sample into the appropriate wells.
- Connect wiring and run gel electrophoresis for 1 hour.
- Analyze gel in UV light and see if samples match the expected size using the ladder as a guide.
