Swartz:Protocols/Autoradiogram

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

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.