Protocol for today:
Used a super sized gel mold and wells. So it took a pretty big volume.
500 ml 1X Low TAE
6.5 g general purpose agarose
Microwave solution for 3 minutes at a time until boiling.
Visually inspect for dense materials (apparently I didn't do well enough as one corner of the gel was higher density than the others.
Placed two rows of 24 well combs in gel mold.
Slowly poured gel into one corner of the mold.
Allowed get to set for 30 minutes.
Once gel was firm I placed it in the electrophoresis chamber.
I filled the chamber with 1 X Low TAE solution until it was even with the gels top surface.
Pipetted 30 ul of Low TAE into each well. Then loaded wells with DNA using the 12 channel pipette in a left to right, top to bottom pattern.
Ran gel for 10 minutes at 120 v.
Paused gel and added 1 ul 5X loading dye to the left most well on both rows.
Ran gel for another 10 minutes at 120 v.
Paused gel and added 1X Low TAE until gel was thoroughly covered.
Ran gel for 1.5 hours until dye had moved appropriately far down the lane.
Placed gel in EtBr bath made 1 L water and 1 ml EtBr.
Soaked gel for 30 minutes.
Imaged on the translinker.
|Top Row||1N1-4 1||1N1-4 2||1N1-4 3||1N1-4 4||1N1-4 5||1N1-4 6||1N1-4 7||1N1-4 8||1N1-4 11||1N1-4 10||1H1-4 1||1H1-4 2||1H1-4 3||1H1-4 4||1H1-4 5||1H1-4 6||1H1-4 7||1H1-4 8||1H1-4 9||1H1-4 10||1S1-4 1||1S1-4 2||1S1-4 3||1S1-4 4|
|Bottom Row||1S1-4 5||1S1-4 6||1S1-4 7||1S1-4 8||1S1-4 9||1S1-4 10||2N1-4 1||2N1-4 2||2N1-4 3||2N1-4 4||2N1-4 5||2N1-4 6||2N1-4 7||2N1-4 8||2N1-4 9||2N1-4 10||2H1-4 1||2H1-4 2||2H1-4 3||2H1-4 4||2H1-4 5||2H1-4 6||2H1-4 7||2H1-4 8|
|Top Row Samples A1-12, B1-12|
|Bottom Row Samples C1-12, D1-12|