03 - Chick Check: The Genetics Behind Blue Chicken Eggs
Module II: Agarose Gel Electrophoresis, continued
POUR 1X Diluted Buffer
REMINDER: Before loading the samples, make sure the gel is properly oriented in the ap- paratus chamber.
RUNNING THE GEL 8.
GEL LOADING TABLE PLACE the gel (on the tray) into an electrophoresis chamber. COVER the gel with 1X TBE electrophoresis buffer (See Table B for recom - mended volumes). The gel should be completely submerged. 9. Using the GEL LOADING TABLE as a guide, LOAD the entire sample (25 µL) into the wells in consecutive order. 10. CHECK that the gel is properly oriented, then PLACE the safety cover onto the chamber. Remember, the DNA samples will migrate toward the positive (red) electrode. 11. CONNECT the leads to the power source and PERFORM electrophoresis (See Table C for time and voltage guidelines). 12. After electrophoresis is complete, REMOVE the gel and casting tray from the electropho- resis chamber. VISUALIZING the SYBR® GEL
LANE 1 2 3 4 5 6
SAMPLE NAME Homozygous non-blue control Homozygous blue control
SLIDE gel off the casting tray onto the viewing surface of the transilluminator. TURN the unit on. DNA should appear as bright green bands on a dark background. PHOTOGRAPH results.
Chicken 1 Chicken 2 Chicken 3 Chicken 4
Time and Voltage Guidelines (2.0% Agarose Gel)
1x TBE Electrophoresis Buffer (Chamber Buffer)
Electrophoresis Model
EDGE™ (10 x 7 cm gel)
M12 (14 x 7 cm gel)
Volts
Min/Max (minutes)
Min/Max (minutes)
150 100
20/30 35/45
30/40 45/55 60/70
EDGE™
150 mL
M12
400 mL
75
NA
35
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