Hosuk:LabNotes/2013-6-25
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Rolony, 11th : Result[edit]
- Done at 06/23
Result[edit]
- SUCCESS!!
- Number of rolonies are much more than previous experiments and their density is so high.
- EM gain need to be decreased because of so much high contrast.
- 10uL Phi29 looked more brighter than 3uL sample.
- EM gain for 10uL is 5, and 10 for 3uL.
- But 3uL of Phi29 is enough.
- Summary of the differences from previous before going to Boston
- Adjust each concentration of Riboshredder+RNase H+RNase H Buffer (Jonathan's protocol)
- Use original CircLigaseII master mix (2uL of enzyme, no doubling other reagents), 4hr CircLigase reaction
- Enzymatics phi29, high concentration (I used 1uL previously and I tried 3uL and 10uL this time, for testing)
- pre-annealing RCA primer in 2x SSC+30% formamide
- and this time I used new vial of aminoallyl-dUTP, I though previous one was almost running out and too old.
Next Plan[edit]
- Compare between…
- Epicentre vs. Enzymatics Phi29
- 1uL vs. 3uL Phi29
- 2hr. vs. 4hr. CircLigase II
- Strip and re-hybridize Cy5 probe
Epi, 20x, NA=0.8, S3(3uL Phi29)[edit]
File:S3 3uLPhi29Enzymat 20x-NA0.8 pos02.png