Matt:LabNotes/2014-2-21

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Screening 32 wells for Ampligase Positive Control Test[edit]

  • Used microscope to check cell density and morphology in each well to pick the best wells for this experiment
    • Color coded each well based on quality of cells
    • Designated 2 or 4 wells for each sample group
      • All wells will have 0.5 U Ampligase/ul except for 2 Fixed Template wells

File:32wellSamples.JPG

Mark & Find Pos -> Well[edit]

  • E6 : Pos 1
  • F6 : Pos 2
  • F7 : Pos 3
  • F8 : Pos 4
  • F9 : Pos 5
  • F10 : Pos 6
  • C7 : Pos 7
  • C8 : Pos 8
  • C9 : Pos 9
  • C10 : Pos 10
  • D7 : Pos 11
  • E7 : Pos 12

Checking MALAT1 Targets in 1000nt RT_RAB7A[edit]

  • In wells: F9 and F10
  • Check with 1uM MALAT1_1stRolonyFISH (Cy3) dye-probe

Checking for non-specific binding of dcProbe2RevComp-Cy3[edit]

  • In wells: F7 and F8
  • Check with 1uM dcProbe2RevComp-Cy3 (ATTO488) dye-probe

Result[edit]

  • No signal -> Dye doesn't have non-specific binding to primary rolonies

Checking MALAT1, RAB7A, and ACTB Targets in RandomHex Primary Rolonies[edit]

  • In wells: E6 and F6
  • Check with 1uM:
    • MALAT1_1stRolonyFISH (Cy3) dye-probe
    • RAB7A_1stRolonyFISH (ATTO550) dye-probe
    • ACTB_1stRolonyFISH (ATTO488) dye-probe