Gel Electrophoresis Protocol

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Gel Electrophoresis Protocol

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