Hosuk:LabNotes/2015-2-6

From ZhangLabWiki
Jump to navigation Jump to search

Human Brain Tissue : CA12k_Nov2014 V4 & V7 : 1st Try[edit]

4th day[edit]


  • 02/06
    1. For V4 & V7 samples: Wash with PBS once
    2. Prepare 4uL of BS(PEG)9 in 196uL PBS
    3. add 200uL BS(PEG)9 mix, incubate for 1hr at RT
    4. Wash with PBS once, add Tris pH 8.0, incubate for 30min
    5. Wash with PBS once


Imaging[edit]

  • For 1st Rolony samples
    1. Prepare detection Probe : Final Conc. 1uM --> 1uL of 100uM Cy3-Adapt + 99uL (2xSSC + 30% Formamide)
    2. Preheat detection Probe at 75C for 5min
    3. Add to sample and incubate at RT for 10min
    4. Wash two times with 2x SSC


Result[edit]

  • Not good!
    • S3: 1st Rolony sample looks fair cell condition, but it has few signal
    • Other 2 dishes (V4 and V7) got damaged…
    • I’ll try more batch!