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Weigh out 0.5g of Agarose on a scale. Pour the the agarose into a 250mL erlenmeyer flask. Pipette 5mL of 5X Agarose Gel Buffer into a 100mL graduated cylinder and dilute 10 fold to a total volume of 50mL. Pour diluted buffer into the agarose erlenmeyer flask and microwave until the solution begins to boil, stopping the microwave and swirling the contents of the flask every 15 seconds. | Weigh out 0.5g of Agarose on a scale. Pour the the agarose into a 250mL erlenmeyer flask. Pipette 5mL of 5X Agarose Gel Buffer into a 100mL graduated cylinder and dilute 10 fold to a total volume of 50mL. Pour diluted buffer into the agarose erlenmeyer flask and microwave until the solution begins to boil, stopping the microwave and swirling the contents of the flask every 15 seconds. | ||
Revision as of 13:08, 23 July 2013
Boston University
BIOMOD 2013 Design Competition
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Protocols
Gel Electrophoresis
Gel Prep:
(For a 1% Agarose Gel)
Weigh out 0.5g of Agarose on a scale. Pour the the agarose into a 250mL erlenmeyer flask. Pipette 5mL of 5X Agarose Gel Buffer into a 100mL graduated cylinder and dilute 10 fold to a total volume of 50mL. Pour diluted buffer into the agarose erlenmeyer flask and microwave until the solution begins to boil, stopping the microwave and swirling the contents of the flask every 15 seconds.
Add 500ul of MgCl2 and 5ul SybrSafe gel dye. Gently swirl the flask to mix contents and then pour the contents into a gel mold and freeze for 20 minutes.
Gel Buffer
To prepare the buffer that fills the gel electrophoresis chamber, pour 30mL of the 5X buffer into a 500mL flask and dilute the total volume of the solution to 300mL. Set aside until the gel is solidified.
Sample Prep
Aliquot the desired volume of sample to be run in the gel, ensuring not to exceed the well capacity. Add loading dye and mix via pipetting.
