Daniel:Notebook/PosSequencing/2017-4-27: Difference between revisions
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Calculations for dye binding: | Calculations for dye binding: | ||
50 × 10<sup>-3</sup> × 500pmol⁄mg = 25 pmol | 50 × 10<sup>-3</sup> mg beads × <sup>500pmol</sup>⁄<sub>mg</sub> = 25 pmol available template @ max | ||
Revision as of 16:00, 27 April 2017
Emulsion PCR (Started Yesterday)
Protocol Part 3-Hybridization and Imaging
Probe hybridization and washes based on Matt's experiment.
Calculations for dye binding:
50 × 10-3 mg beads × 500pmol⁄mg = 25 pmol available template @ max
- Fluorescent Oligo Binding
- Put suspension on magnet
- Resuspend beads in 25 uL 4X SSC buffer
- Add 25 uL fluorescent probe in 60% formamide
- Incubate at room temp in the dark for 30 min
- Wash three times in 2X SSC
- Imaging