Daniel:Notebook/PosSequencing/2017-7-10

From ZhangLabWiki
Revision as of 20:12, 10 July 2017 by >Djacobse (Created page with "=emPCR8 (Started Thursday July 6)= Back to Calendar ==Dye Hybridization and Imaging== Since D...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

emPCR8 (Started Thursday July 6)

Back to Calendar

Dye Hybridization and Imaging

Since Saturday's results showed nothing, I'm going to try again but with the protocol the same as the first Lambda exonuclease test

  1. Dye hybridization
    1. Mix 20 uL ssDNA (lambda digested) and 20 uL 2X SSC
    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