Team:GeorgiaTech/Gel Electrophoresis Protocol

From 2013.igem.org

Procedure

  • Get a agarose gel from the refrigerator and place in apparatus, making sure it is completely covered in TAE buffer.
  • Calculate the amount of purified plasmid that needs to be added to each lane.
    • The goal is to add ~100ng of DNA to each well.
  • The typical ladders used are 1k bp from NEB (only 2uL is used)
  • In a PCR tube mix:
    • desired amount (100ng) of DNA
    • 2uL of 6x dye
    • 2 uL of Buffer2
    • 3 uL of water
    • 0.5 uL of each restriction enzyme
  • Load 8uL into each lane on the gel. Run the gel for 30min at default settings
  • Look at the gel under blue lamp and take a picture
  • Discard the gel and clean up