04 - Put the M into STEM: Quantitative Techniques for Biotechnology
Step 1: Agarose Gel Electrophoresis
RUNNING THE GEL
8. PLACE the gel (still 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.
9. PUNCTURE the foil overlay of the QuickStrip™ with a pipet tip. LOAD the entire sample (35 µL) into the well in the order indicated by Table 1, at right. 10. PLACE safety cover on the unit. CHECK that the gel is properly ori- ented. Remember, the DNA samples will migrate toward the posi- tive (red) electrode. 11. CONNECT leads to the power source and
REMINDER: Before loading the samples, make sure the gel is properly oriented in the apparatus chamber.
PERFORM electrophoresis (See Table C for time and voltage guidelines). Allow the tracking dye to migrate at least 3 cm from the wells. 12. After electrophoresis is complete, RE- MOVE the gel and casting tray from the electrophoresis chamber and PROCEED to gel staining.
Tube A Tube B Tube C Tube D Tube E Tube F TABLE 1: GEL LOADING 10 cycles 20 cycles 30 cycles
Lane 1 2 3 4 5 6
DNA Standard Marker 0 cycles 5 cycles
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
45
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