Hosuk:LabNotes/2014-2-9
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NCIH1975 : Mutation Detection : 2nd Try, Data Analysis
- Data from 2/3
- Took pictures at 3 Positions on the sample in MatTek dish
Procedure
- Maximum projection image with all images of 8 steps
- Run PISA to make BW images
- Use the BW image as mask image file and do masking an each image (=image file .* mask image)
- Run PISA with masked image, and obtain a number of rolonies, centroid coordinate information of each rolony
- Decode the rolony to extract decoding information along steps
Major Parameters
Decode Binary table of each target
EGFR_T790M_WT | EGFR_T790M_MU | EGFR_L858R_WT | EGFR_L858R_MU | |
Step1 (1FAM) | 0 | 1 | 0 | 1 |
Step2 (1Cy3) | 1 | 0 | 0 | 0 |
Step3 (2FAM) | 0 | 0 | 1 | 0 |
Step4 (2Cy3) | 0 | 0 | 0 | 1 |
Step5 (3FAM) | 1 | 0 | 1 | 0 |
Step6 (3Cy3) | 0 | 1 | 0 | 0 |
Step7 (4FAM) | 1 | 0 | 0 | 1 |
Step8 (4Cy3) | 0 | 1 | 1 | 0 |
Decode Binary String of each target
- EGFR_T790M_WT = '01010010'
- EGFR_T790M_MU = '10100001'
- EGFR_L858R_WT = '10010100'
- EGFR_L858R_MU = '01001001'
1 bit Error cases of Decode Binary String
- EGFR_T790M_WT
- Normal : '01010010'
- Error 1 : '01010000'
- Error 2 : '01000010'
- Error 3 : '00010010'
- EGFR_T790M_MU
- Normal : '10100001'
- Error 1 : '10100000'
- Error 2 : '10000001'
- Error 3 : '00100001'
- EGFR_L858R_WT
- Normal : '10010100'
- Error 1 : '10010000'
- Error 2 : '10000100'
- Error 3 : '00010100'
- EGFR_L858R_MU
- Normal : '01001001'
- Error 1 : '01001000'
- Error 2 : '01000001'
- Error 3 : '00001001'