AlanFung:LabNotes/Rolony/2013-9-26: Difference between revisions
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>Alan6017518 |
>Alan6017518 (→Goal) |
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*Call a base based on the signal area | *Call a base based on the signal area | ||
*Generate the 3 base sequence based on the location of the signal | *Generate the 3 base sequence based on the location of the signal | ||
==Tools== | |||
*Images (taken with Hosuk) | |||
*Matlab | |||
==Protocol== |
Revision as of 21:47, 26 September 2013
Strip off dye coupling with formamide
- Aspirate PBS from mattek dish with primary/secondary rolonies
- Add 200ul preheated (65C-80C) 80% formamide in 2X SSC
- Incubate 10min RT
- Rinse w H2O
- Add in 1:200-1uM 200ul (preheat 65C) ACTB-primary rolony target probe (Alexa 488) (200uM ACTB_1stRolonyFISH)
- Incubate 10min RT
- Rinse w 2x SSC twice
- Add in 2X ssc
- image
Matlab Image Analysis
Goal
- The goal is to analyze the image taken from SBS sequencing of the primary rolony
- I would create a threshold to screen out the noise
- Call a base based on the signal area
- Generate the 3 base sequence based on the location of the signal
Tools
- Images (taken with Hosuk)
- Matlab