Afra

LOADING

1. Fill the running chamber with TAE+EtBr solution
2. Take the tape off from the gel tray when the gel is totally set
3. Place the gel/gel tray into the chamber and top up with some more of TAE+EtBr solution (approx. 1 cm above the gel)
4. Remove the comb
5. Add 5 uL of Orange-G (dye) into samples – aliquotes found in Fridge #1, first draw
6. Load 1.5 uL of DNA ladder (0.25 ug/uL; invitrogen) in the first well – in Fridge #1 shelf
7. Load 12.5 uL of sample (half of total PCR volume) into each well using a multi-pipette
8. Load 1.5 uL of DNA ladder in the last well
9. Connect the chamber to the battery and run until you get a good separation

Battery Settings

• Voltage select: set at desired voltage depending on how fast you want to run the gel
• Range select: Low
• Choose volts (not mA)
• ALWAYS have the black end (-ve end) closer to the wells

DEVELOPING

1. Take the gel into the UV room on a tray (always wear gloves!)
2. Place the gel into the bottom draw of the Gel doc (Bio Rad) machine
3. On computer, select: Quantity One – File – Gel doc
4. Select “Manual exposure”, then press “Trans UV” on the Gel doc machine to view DNA bands
5. Enlarge the view field by “zoom” button & Optimise by “low/high” scale on “display”
6. Print the figure by pressing “Video print”

Solutions: