AlanFung:LabNotes/Rolony/2013-9-26: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Alan6017518
>Alan6017518
Line 15: Line 15:
*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

Protocol