Haynes Lab:Notebook/Engineering PC-TFs/2012/10/16: Difference between revisions
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==Summary== | ==Summary== | ||
Gel Electrophoresis | Gel Electrophoresis | ||
Follow steps for gel electrophoresis. | *Follow steps for gel electrophoresis. | ||
Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. | *Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. | ||
Create 1% gel by putting .6 grams of agarose into flask. | *Create 1% gel by putting .6 grams of agarose into flask. | ||
Microwave agarose solution for 40 seconds | *Microwave agarose solution for 40 seconds | ||
Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds. | *Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds. | ||
When flask is taken out of microwave, make sure that the agarose is completed dissolved. | *When flask is taken out of microwave, make sure that the agarose is completed dissolved. | ||
Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the large "teeth" and slip in the notches. | *Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the large "teeth" and slip in the notches. | ||
Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL. | *Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL. | ||
Pour gel into tray. Wait twenty minutes for gel to settle. | *Pour gel into tray. Wait twenty minutes for gel to settle. | ||
Wash the agarose gel flask. | *Wash the agarose gel flask. | ||
Pipette 15 μL of DNA ladder into first well. Pipette 45μL of PCR reactions into subsequent wells. | *Pipette 15 μL of DNA ladder into first well. Pipette 45μL of PCR reactions into subsequent wells. | ||
Turn on electrophoresis and let run for thirty minutes. | *Turn on electrophoresis and let run for thirty minutes. | ||
Revision as of 14:52, 19 October 2012
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SummaryGel Electrophoresis
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