Genotyping- PCR
PCR- Gel Electrophoresis/ Running the Gel(Beads and Green Mix)
1. Pour 800ml of 1x TAE Buffer into gel chamber
2. Position gel to allow the DNA to run from BlackRed (Cathode Anode)
3. For bead PCRs: Add 3µL Orange G using yellow tips to each PCR tube and eject
the 1-200µL yellow tips into each tube to conserve tips. Mix Orange G and PCR
samples by pipetting up and down before aspirating to load the specimen into the
appropriate well, then load 10µL of PCR sample to an appropriate well on gel .
Orange G will migrate.
4. For green mix PCRs: Load 10µL of each PCR sample to an appropriate well on
the gel. No Orange G required! Green mix will separate into blue(slow) and
yellow (fast) band
5. Add 10µL of DNA ladder (Fisher: BP2573100) into well #1 of each row of wells.
6. Attach the lid of the chamber and insert the corresponding red and black plugs
7. Turn the machine on (~ 130 amps) and run the gel for ~1-1½ hrs or until the
orange band/yellow band has migrated to a desired location on the gel
* Once the orange band/yellow band has migrated to an end point, Turn the machine off
and remove the lid. Gel can then be removed to take pictures. Camera is on the 10 th floor.
Gel is disposed of in the Ethidium Bromide Hazardous Waste container on the 10th floor.
Written by Shan; Updated by LM on 4/21/15
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