Hosuk:LabNotes/2014-1-27: Difference between revisions

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*Raw: [[Media:MIP_NCIH1975_Mutant_Pos1_Step0_Cy5.tif|Pos1]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step0_Cy5.tif|Pos2]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step0_Cy5.tif|Pos3]]
*Raw: [[Media:MIP_NCIH1975_Mutant_Pos1_Step0_Cy5.tif|Pos1]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step0_Cy5.tif|Pos2]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step0_Cy5.tif|Pos3]]
[[File:NCIH1975_Mutant_Pos1-2-3_1stRolony.png|900px]]
[[File:NCIH1975_Mutant_Pos1-2-3_1stRolony.png|1000px]]




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*'''Pos1''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos1_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
*'''Pos1''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos1_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos1_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
[[File:NCIH1975_Mutant_Pos1_dcProbe.png|600px]]
[[File:NCIH1975_Mutant_Pos1_dcProbe.png|800px]]




*'''Pos2''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos2_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
*'''Pos2''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos2_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos2_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
[[File:NCIH1975_Mutant_Pos2_dcProbe.png|600px]]
[[File:NCIH1975_Mutant_Pos2_dcProbe.png|800px]]




*'''Pos3''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos3_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
*'''Pos3''' --> Raw : [[Media:MIP_NCIH1975_Mutant_Pos3_Step1_dcProbe1-Cy3_edit_Mark.tif|dcProbe1]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step2_dcProbe2-Cy3_edit_Mark.tif|dcProbe2]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step3_dcProbe3-Cy3_edit_Mark.tif|dcProbe3]], [[Media:MIP_NCIH1975_Mutant_Pos3_Step4_dcProbe4-Cy3_edit_Mark.tif|dcProbe4]]
[[File:NCIH1975_Mutant_Pos3_dcProbe.png|600px]]
[[File:NCIH1975_Mutant_Pos3_dcProbe.png|800px]]

Latest revision as of 02:06, 28 January 2014

NCIH1975 : Mutation Detection[edit]


2nd Rolony[edit]

  • Annealing Padlock probes, Ampligase, and 2nd RCA fab. --> done at 01/19~01/21


Detection of Mutation[edit]

  • Procedure
    1. Add Cy5-Adapt to detect 1st Rolonies, 3 positions
    2. Strip (80% Formamide for 15min), Add dcProbe1 (incubate 15min), image
    3. Strip (80% Formamide for 15min), Add dcProbe2 (incubate 15min), image
    4. Strip (80% Formamide for 15min), Add dcProbe3 (incubate 15min), image
    5. Strip (80% Formamide for 15min), Add dcProbe4 (incubate 15min), image


Result[edit]

  • Image at 3 different positions


1st Rolony[edit]
  • Pos1 : 5151
  • Pos2 : 5964
  • Pos3 : 6485

File:NCIH1975 Mutant Pos1-2-3 1stRolony.png


Each dcPobes[edit]
  • Not many, just a few, 10~20 from 2nd rolonies.
  • Need positive control or some information about expected abundance of each gene.

File:NCIH1975 Mutant dcProbeCount.png


  • Images of each gene at each position
    • Circles are the 2nd Rolony that I've counted.


File:NCIH1975 Mutant Pos1 dcProbe.png


File:NCIH1975 Mutant Pos2 dcProbe.png


File:NCIH1975 Mutant Pos3 dcProbe.png