Hosuk:LabNotes/2014-3-26

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42C RT incubation time and New CircLigase II Reaction Buffer Test[edit]

  • Test new, home made CircLigase II buffer in fixed cell sample
  • And 42C RT reaction temperature test


  • New reaction buffer is used for all samples.
    1. Dish1 : PGP1F, random hexamer, 37C RT temp.
    2. Dish2 : PGP1F, RAB7A specific RT primer, 37C RT temp.
    3. Dish3 : PGP1F, random hexamer, 42C RT temp.
    4. Dish4 : PGP1F, RAB7A specific RT primer, 42C RT temp.


Result[edit]

  • RT at 42C incubation works well
  • New CircLigase II Buffer works well
  • Position specific RT primer of RAB7A (220bp) makes many rolonies.
  • And there are a few of RAB7A target detected, I’m currently analyzing how many RAB7A targets are there.
  • Images with 63x have more Rolonies/cell, I think the reason is 63x has smaller field of view, so there are much fewer cells in a field of view, but 63x obj. can detect many rolony signals.
  • RAB7A targets are about ~5% out of 1st rolonies, which is always found in other experiments.

File:RolonyCount 20140326 with3D.png