Daniel:Notebook/PosSequencing/2017-5-22

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Single Stranded Probe Binding Test (Started Friday May 19)

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Protocol (Continued from Saturday May 20)

  1. Dye hybridization
    1. Mix 20 uL ssDNA (lambda digested) and 20 uL dsDNA (original, non-digested template)
    2. Add 1 uL appropriate (10 uM) dye, A488-dc6 for sample 1 and Cy5-dc6 for sample 3
    3. Incubate in the dark (RT) for 15 minutes
    4. Magnet pulldown, remove supernatant and add 40 uL 2X SSC buffer
    5. Add 15 uL to a slide for analysis
    6. Seal coverslip with nail polish and wait approximately 30 minutes to dry
    7. Image on microscope