Haynes Lab:Notebook/Engineering PC-TFs/2012/10/02: Difference between revisions
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==Summary== | ==Summary== | ||
* Gel Electrophoresis | * Gel Electrophoresis | ||
Follow steps for gel electrophoresis. | |||
Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. | |||
Fill gel flask with up to 60 ml of TA buffer. | |||
Create 1% gel by putting .6 grams of agarose into flask. | |||
Microwave agarose solution for 30 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. | |||
Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "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. | |||
Pour gel into tray. Wait twenty minutes for gel to settle. | |||
Wash the agarose gel flask. | |||
Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells. | |||
Turn on electrophoresis and let run for thirty minutes. | |||
Revision as of 09:52, 3 October 2012
 Engineering PC-TFs
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Summary
Follow steps for gel electrophoresis. Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray. Fill gel flask with up to 60 ml of TA buffer. Create 1% gel by putting .6 grams of agarose into flask. Microwave agarose solution for 30 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. Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "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. Pour gel into tray. Wait twenty minutes for gel to settle. Wash the agarose gel flask. Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells. Turn on electrophoresis and let run for thirty minutes.
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