Hosuk:LabNotes/2013-6-25

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Revision as of 16:04, 26 June 2013 by >Hosuki78 (Created page with "*LabNote ===Rolony, 11th : Result=== *Done at 06/23 ====Result==== *'''SUCCESS!!''' *Number of rolonies are much more than...")
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Rolony, 11th : Result[edit]

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…
    1. Epicentre vs. Enzymatics Phi29
    2. 1uL vs. 3uL Phi29
    3. 2hr. vs. 4hr. CircLigase II
    4. Strip and re-hybridize Cy5 probe



Epi, 20x, NA=0.8, S3(3uL Phi29)[edit]

File:S3 3uLPhi29Enzymat 20x-NA0.8 pos02.png


Epi, 20x, NA=0.8, S4(10uL Phi29)[edit]

File:S4 10uLPhi29Enzymat 20x-NA0.8 pos03.png