Team:CSU Fort Collins/Notebook/Protocols=Gel

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.