Team:CSU Fort Collins/Notebook/Protocols=Gel
From 2014.igem.org
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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> |
Revision as of 18:25, 9 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.