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===NCIH1975 : Mutation Detection=== *Continued from [[Hosuk:LabNotes/2014-1-19|1/19]] ====2nd Rolony==== *Annealing Padlock probes, Ampligase, and 2nd RCA fab. --> done at 01/19~01/21 ====Detection of Mutation==== *Procedure *#Add Cy5-Adapt to detect 1st Rolonies, 3 positions *#Strip (80% Formamide for 15min), Add dcProbe1 (incubate 15min), image *#Strip (80% Formamide for 15min), Add dcProbe2 (incubate 15min), image *#Strip (80% Formamide for 15min), Add dcProbe3 (incubate 15min), image *#Strip (80% Formamide for 15min), Add dcProbe4 (incubate 15min), image ====Result==== *Image at 3 different positions =====1st Rolony===== *Pos1 : 5151 *Pos2 : 5964 *Pos3 : 6485 *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|1000px]] =====Each dcPobes===== *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|500px]] *Images of each gene at each position **Circles are the 2nd Rolony that I've counted. *'''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|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]] [[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]] [[File:NCIH1975_Mutant_Pos3_dcProbe.png|800px]]
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