07 - Lion Family Reunion: Conservation Biology Genetics
Agarose Gel Electrophoresis, continued
9. PLACE the gel (on the tray) into the electrophoresis chamber. COVER the gel with 1X electrophoresis buffer (See Table B for recommended volumes). The gel should be completely submerged. 10. LOAD the entire sample (35 µL) into the well in the order indicated by Table 1. 11. 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. 12. CONNECT the leads to the power source and PER-
FORM electrophoresis (See Table C for time and voltage guidelines). complete, REMOVE the gel and casting tray from the electrophoresis chamber. PROCEED TO VISUALIZATION. 13. After electrophoresis is
Table 1: Gel Loading
Lane 1 2 3 4 5
Tube
Sample
Tube A
Standard DNA Marker
Tube B
Lion A’s mtDNA cut with Enzyme 1
Tube C
Lion A’s mtDNA cut with Enzyme 2
Tube D
Lion B’s mtDNA cut with Enzyme 1
Tube E
Lion B’s mtDNA cut with Enzyme 2
Time and Voltage Guidelines (0.8% Agarose Gel)
1x Electrophoresis Buffer (Chamber Buffer)
EDGE™
M12 & M36
Volts
Min/Max (minutes)
Min/Max (minutes)
EDGE™
150 mL
3 mL
147 mL
150 125 100
10/20
20/35 30/45 40/60
M12
400 mL
8 mL
392 mL
N/A
M36
1000 mL
20 mL
980 mL
15/25
83
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