04 - Follow That Flush: Using Biotechnology for Early Disease Detection
Module II: Agarose Gel Electrophoresis, continued
REMINDER: Before loading the samples, make sure the gel is properly oriented in the
RUNNING THE GEL 9.
PLACE gel (on the tray) into electrophoresis chamber. COVER the gel with 1X electrophoresis buffer (See Table B for recommended vol - umes). The gel should be completely submerged.
apparatus chamber.
10. PUNCTURE the foil overlay of the QuickStrip™ with a pipet tip. LOAD the entire sample (35 µL) into the well as indicated by the Gel Loading table. 11. PLACE safety cover. CHECK that the gel is properly oriented. Remember, the DNA samples will migrate toward the positive (red) electrode. 12. CONNECT leads to the power source and PERFORM electrophoresis. (See Table C for time and voltage guidelines.) 13. After electrophoresis is complete,
REMOVE the gel and casting tray from the electrophoresis chamber.
GEL LOADING TABLE
Lane 1 2 3 4 5 6
Tube A Tube B Tube C Tube D Tube E Tube F
DNA Standard Marker Sample 1 - Treatment Plant Sample 2 - Community Center Sample 3 - University Dorms Sample 4 - Downtown Sample 5 - Regional Airport
VISUALIZING the SYBR® GEL 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 back- ground. PHOTOGRAPH results.
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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