Daniel:Notebook/PosSequencing/2017-7-14: Difference between revisions
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>Djacobse (Created page with "=emPCR 9-Concentration Comparison (Started Wednesday)= Daniel:Notebook/PosSequencing|Back to Ca...") |
>Djacobse |
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<li>Dye hybridization</li> | <li>Dye hybridization</li> | ||
<ol type="A"> | <ol type="A"> | ||
<li> | <li>Resuspend beads in 30 uL 2X SSC</li> | ||
<li>Incubate in the dark at | <li>Mix 15 uL 4X SSC, 9 uL nfH<sub>2</sub>O, 3 uL 10 uM dcprobe6-A488, and 3 uL 10 uM dcprobe6-Cy5</li> | ||
<li> | <li>Incubate master mix in the dark at 80C for 5 minutes</li> | ||
<li>Incubate in the dark | <li>Add 10 uL master mix to each sample</li> | ||
<li>Incubate at 55C for 30 minutes in the dark</li> | |||
<li>Magnet pull down, remove supernatant and add 40 uL 2X SSC buffer</li> | <li>Magnet pull down, remove supernatant and add 40 uL 2X SSC buffer</li> | ||
<li>Add 15 uL to a slide for analysis</li> | <li>Add 15 uL to a slide for analysis</li> |
Revision as of 17:56, 14 July 2017
emPCR 9-Concentration Comparison (Started Wednesday)
Protocol-Part 3
- Dye hybridization
- Resuspend beads in 30 uL 2X SSC
- Mix 15 uL 4X SSC, 9 uL nfH2O, 3 uL 10 uM dcprobe6-A488, and 3 uL 10 uM dcprobe6-Cy5
- Incubate master mix in the dark at 80C for 5 minutes
- Add 10 uL master mix to each sample
- Incubate at 55C for 30 minutes in the dark
- Magnet pull down, remove supernatant and add 40 uL 2X SSC buffer
- Add 15 uL to a slide for analysis
- Seal coverslip with nail polish and wait approximately 30 minutes to dry