Swartz:Protocols/Autoradiogram

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https://docs.google.com/document/d/1vgVufOvnKvn84drFBmOtEqhZQEKNFzKqw82ItypqvHI/edit

Contents

How to do an Autoradiogram

Kurt Knapp
10/25/04
Uploaded by Chris VanLang 9/21/11

Drying the Gel

  • Rinse the gel in water once
  • Lay it on a piece of filter paper
  • Put it under the plastic sheet of the gel dryer
    • Lay a piece of saran wrap over the top of the gel.
  • Turn on the coldtrap and gel dryer
  • Close the valve on the coldtrap
  • Turn on the house vacuum
  • Check that the vacuum toggle switch is on the gel dryer
  • Press start on the gel dryer
  • Stop the gel after 1hour … it should be dry

Setting up Cassette

  • Blank a phosphor screen cassette using the light box for 20 minutes.
  • On the gel, use a pen to mark the protein ladder.
  • Use 1:10 mixture of C14 Leucine and spot the pen marks.
  • Tape the gel against the felt of the cassette so it doesn’t move.
    • It is best to orient the gel so that the ladder is on the left starting from B2.
  • Store for 1-2 days

Developing Cassette

  • Use the imager from the Clark center.
  • Email Anja Duursma via aduursma@stanford.edu to get access. You will need your sunetID.
  • Remember to bring your own USB drive as you cannot store any data on the computer.
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