Difference: DenaturingFormaldehydeGels (4 vs. 5)

Revision 52018-10-26 - SimonDAlton

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META TOPICPARENT name="ProtocolList"

Denaturing Formaldehyde Gels for Verifying RNA size

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  1. Remove the gel from the rig when the blue loading dye is approximately 1/2-3/4 down the gel. If the predicted RNA size is 4kb or larger, leave gel til 3/4 to get good resolution, smaller RNAs can be left for the shorter time.
  2. Wash quickly once in 1x TAE, followed by a longer, 20 minute wash in 1x TAE. This will bring the pH to a more neutral value to allow Sybrgold to bind to RNA.
  3. Incubate gel in 1x Sybrgold (from 10k x stock) in 1x TAE for 30 mins.
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  1. Image as usual.
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  1. Image as usual. If signal to background is low, destain gel in 1xTAE for 15 mins and image again.
 

Bonus Feature - Preparation of DEPC water

 
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