Brown: Team Resistance/21 June 2008


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21 June 2008

Gel Electrophoresis

  • Gel Electrophoresis is primarily used to determine the length of a fragment of DNA. DNA Ladders are run beside experimental fragments as points of reference. A DNA Ladder contains all different lengths of DNA that when run out on a gel, separate to reveal the number of base pairs in each gene.
  1. Prepare the casting gel: 1% agarose gel - 1) 0.5 g agarose 2) 50 mL TBE + SYBR safe.
  2. Gel must be submerged completely with 300-500 mL (large gel tray) .5X TBE + SYBR safe buffer.
  3. Put the gel, WITHOUT the lid, in the microwave on HIGH for 1 minute.
  4. Cool the bottle of gel under water.
  5. Pour into the castings with rubber stoppers.
  6. The gel takes approximately 20 minutes to set. Once gel is set make sure to remove the rubber stopprs.
  7. DNA has a negative charge. The wells of the gel are aligned with on the negative side (Cathode). DNA will move away from the Cathode to the Anode (+).
  • The gel acts as a sieve for fragements as small as 100 base pairs and as large as 50,000 base pairs. 0.3% large fragments - 2% large fragments.
  1. When complete, pour TBE buffer back into a container. The buffer can be used up to 8 times.