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.
- Dish1 : PGP1F, random hexamer, 37C RT temp.
- Dish2 : PGP1F, RAB7A specific RT primer, 37C RT temp.
- Dish3 : PGP1F, random hexamer, 42C RT temp.
- 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.