Dye/Indicator Lab Flow Chart - authors@Fred Hutch
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group:*SEP Files:*Uber SEP = Index:Kits:*Kits:Gel Kit :3Dye Lab:DyeLabFlowWd_08.doc 6/18/2008 8:49 AM 1 Weigh agarose powder Mix agarose & 1X TAE buffer Microwave to dissolve, swirl occasionally Optional: Keep agarose in 60˚C waterbath Slowly pour agarose into gel tray Place comb in gel. Dye lab=middle slot Let gel harden. Remove comb and dams after pouring thin layer of buffer over the gel. Label 8 microcentrifuge tubes 1-8 Dye/Indicator Lab Flow Chart Set up gel tray in gel box with dams at each end.
Transcript of Dye/Indicator Lab Flow Chart - authors@Fred Hutch
group:*SEP Files:*Uber SEP = Index:Kits:*Kits:Gel Kit :3Dye Lab:DyeLabFlowWd_08.doc 6/18/2008 8:49 AM 1
Weigh agarose powder
Mix agarose & 1X TAE buffer
Microwave to dissolve, swirl occasionally
Optional: Keep agarose in 60˚C waterbath
Slowly pour agarose into gel tray
Place comb in gel. Dye lab=middle slot Let gel harden.
Remove comb and dams after pouring thin layer of buffer over the gel.
Label 8 microcentrifuge tubes 1-8
Dye/Indicator Lab Flow Chart
Set up gel tray in gel box with dams at each end.
group:*SEP Files:*Uber SEP = Index:Kits:*Kits:Gel Kit :3Dye Lab:DyeLabFlowWd_08.doc 6/18/2008 8:49 AM 2
Fill out a diagram with dye names Add 20 µl dye in each tube
Centrifuge tubes
Load gel wells with 15 µl dye each
Connect gel box to power supply, run gel 10 min at 100 V
Turn off power source and disconnect. Lift gel tray out of box, and slide gel onto plastic wrap on the table. Keep track of + and - ends of gel.
Lay acetate over gel. With permanent marker, trace locations of wells, dye spots, and + and - end of gels.
Set gel on blotting paper. Press with weight for about 5 seconds. Repeat to make a blot for each person in the group.
Use acetate trace to measure mm traveled from well by each dye spot.
Clean up and return all materials and equipment.
1. Bromophenol blue
2. Orange G 3. Etc. 4. 5. 6. 7. 8.
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