Brown: Team Resistance/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.