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...") |
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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> | ||
Line 17: | Line 18: | ||
</ol> | </ol> | ||
</ol> | </ol> | ||
==Results== | |||
<gallery perrow=2 heights=300px widths=300px mode=packed-hover caption="10 nM template Concentration"> | |||
File:EmPCR9-20170714.lif_10nM-Field5-SNAP2_ch00.jpg|Field 5 | |||
</gallery> | |||
<gallery perrow=2 heights=300px widths=300px mode=packed-hover caption="100 pM template Concentration"> | |||
File:EmPCR9-20170714.lif_100pM-Field1-SNAP_ch00.jpg|Field 1 | |||
File:EmPCR9-20170714.lif_100pM-Field4-SNAP_ch00.jpg|Field 4 | |||
File:EmPCR9-20170714.lif_100pM-Field6-SNAP_ch00.jpg|Field 6 | |||
File:EmPCR9-20170714.lif_100pM-Field7-SNAP_ch00.jpg|Field 7 | |||
</gallery> | |||
<gallery perrow=2 heights=300px widths=300px mode=packed-hover caption="10 pM template Concentration"> | |||
File:EmPCR9-20170714.lif_10pM-Field1-SNAP_ch00.jpg|Field 1 | |||
File:EmPCR9-20170714.lif_10pM-Field3-SNAP_ch00.jpg|Field 3 | |||
File:EmPCR9-20170714.lif_10pM-Field7-SNAP_ch00.jpg|Field 7 | |||
File:EmPCR9-20170714.lif_10pM-Field9-SNAP_ch00.jpg|Field 9 | |||
</gallery> | |||
[[Category:PosSeq]] [[Category:20170712]] |
Latest revision as of 22:40, 14 July 2017
emPCR 9-Concentration Comparison (Started Wednesday)[edit]
Protocol-Part 3[edit]
- 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
Results[edit]
- 10 nM template Concentration
- EmPCR9-20170714.lif 10nM-Field5-SNAP2 ch00.jpg
Field 5
- 100 pM template Concentration
- EmPCR9-20170714.lif 100pM-Field1-SNAP ch00.jpg
Field 1
- EmPCR9-20170714.lif 100pM-Field4-SNAP ch00.jpg
Field 4
- EmPCR9-20170714.lif 100pM-Field6-SNAP ch00.jpg
Field 6
- EmPCR9-20170714.lif 100pM-Field7-SNAP ch00.jpg
Field 7
- 10 pM template Concentration
- EmPCR9-20170714.lif 10pM-Field1-SNAP ch00.jpg
Field 1
- EmPCR9-20170714.lif 10pM-Field3-SNAP ch00.jpg
Field 3
- EmPCR9-20170714.lif 10pM-Field7-SNAP ch00.jpg
Field 7
- EmPCR9-20170714.lif 10pM-Field9-SNAP ch00.jpg
Field 9