Daniel:Notebook/PosSequencing/2017-4-27: Difference between revisions

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==Protocol Part 3-Hybridization and Imaging==
==Protocol Part 3-Hybridization and Imaging==


Probe hybridization and washes based on [[Matt:LabNotes/2015-3-12|Matt's experiment]]. Calculations for dye binding:
Probe hybridization and washes based on [[Matt:LabNotes/2015-3-12|Matt's experiment]].  


&alpha; 50&times;10<sup>-3</sup>&times;500pmol&frasl;mg
Calculations for dye binding:
 
50 &times; 10<sup>-3</sup> &times; 500pmol&frasl;mg = 25 pmol





Revision as of 15:58, 27 April 2017

Emulsion PCR (Started Yesterday)

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Protocol Part 3-Hybridization and Imaging

Probe hybridization and washes based on Matt's experiment.

Calculations for dye binding:

50 × 10-3 × 500pmol⁄mg = 25 pmol


  1. Fluorescent Oligo Binding
    1. Put suspension on magnet
    2. Resuspend beads in 25 uL 4X SSC buffer
    3. Add 25 uL fluorescent probe in 60% formamide
    4. Incubate at room temp in the dark for 30 min
    5. Wash three times in 2X SSC
  2. Imaging