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

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Calculations for dye binding:  
Calculations for dye binding:  


50 &times; 10<sup>-3</sup> &times; 500pmol&frasl;mg = 25 pmol
50 &times; 10<sup>-3</sup> mg beads &times; <sup>500pmol</sup>&frasl;<sub>mg</sub> = 25 pmol available template @ max





Revision as of 16:00, 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 mg beads × 500pmolmg = 25 pmol available template @ max


  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