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

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(Summary)
(Summary)
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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.
Fill gel flask with up to 60 ml of TA buffer.
+
*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 30 seconds
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.
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 thin "teeth" and slip in the notches.
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.
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 5μL of PCR reactions into subsequent wells.
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.
Turn on electrophoresis and let run for thirty minutes.
 
  
  

Revision as of 08:53, 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.