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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Summary

Gel Electrophoresis

  • Follow steps for gel electrophoresis.
  • Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray.
  • Create 1% gel by putting .6 grams of agarose into flask.
  • 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.
  • 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.
  • 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 45μL of PCR reactions into subsequent wells.
  • Turn on electrophoresis and let run for thirty minutes.



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