Haynes Lab:Notebook/Engineering PC-TFs/2012/10/02: Difference between revisions

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* Gel Electrophoresis
* 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 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.


[[Image:October_2_gel|400px|thumb|left|PCR Check]]
 
 
[[Image:October_2_gel.jpg|400px|thumb|left|PCR Check]]


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Revision as of 09:54, 3 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 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.


PCR Check


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