CHEF Gel Troubleshooting: Help With Plugs & Ladder

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Discussion Overview

The discussion centers around troubleshooting issues related to running a CHEF gel, specifically focusing on problems with plugs not leaving the wells and the presence of a thick band in the well as indicated by ethidium bromide staining. Participants are seeking advice and expertise on the matter.

Discussion Character

  • Technical explanation, Debate/contested

Main Points Raised

  • One participant reports that no plugs have left the wells and mentions the presence of a thick band in the well.
  • Another participant inquires whether any material has exited the well and suggests ruling out power supply issues.
  • A later reply confirms that nothing has exited the well and notes that power supply issues have not yet been ruled out.
  • One participant emphasizes that checking the power supply is a priority and recommends using a voltmeter or an oscilloscope if the power supply applies a pulsed potential.

Areas of Agreement / Disagreement

Participants do not appear to reach a consensus, as there is uncertainty regarding the power supply and its potential role in the issue being discussed.

Contextual Notes

There are limitations regarding the assumptions made about the power supply and the specific conditions under which the gel is being run, which remain unresolved.

MatthewHaas
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Has anyone ever ran a CHEF gel before? I am having trouble getting my plugs to leave the wells. Ethidium bromide staining shows a thick band in the well, and I am supposed to have a ladder. I have some images I can post, but I figured it makes more sense to first see if anyone out there has any expertise in the area.

Thanks!
 
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Did anything get out of the well? I assume that you have already ruled out power supply issues, yes?
 
I am sorry for the late reply. No, nothing got out of the well. We have not rulerd out power supply issues.
 
Power supply is the first thing you check. Use a voltmeter unless the power supply applies a pulsed potential. If it is pulsed, you need an oscilliscope.
 

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