"
Team:CSU Fort Collins/Notebook/Protocols=Gel
From 2014.igem.org
(Difference between revisions)
| Line 687: | Line 687: | ||
<div class='wrapper'> | <div class='wrapper'> | ||
<center> | <center> | ||
| - | |||
<div class="container"> | <div class="container"> | ||
<h3>Gel Electrophoresis Protocol</h3> | <h3>Gel Electrophoresis Protocol</h3> | ||
| + | |||
| + | <ul> | ||
| + | <li>Using a balance, mass out 1.0 gram of agarose. Mix this with 100 MICROL of TAE 1X Buffer.</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>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> | ||
| + | <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 MICROL of each sample into the appropriate wells.</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> | ||
| + | </ul> | ||
| + | |||
Revision as of 06:04, 8 October 2014
Gel Electrophoresis Protocol
- Using a balance, mass out 1.0 gram of agarose. Mix this with 100 MICROL 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 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.
- 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 MICROL 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.
