Daniel:Notebook/PosSequencing/2017-7-12: Difference between revisions
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>Djacobse m (→Protocol) |
>Djacobse |
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<li>Dye hybridization</li> | <li>Dye hybridization</li> | ||
<ol type="A"> | <ol type="A"> | ||
<li>Mix 20 uL ssDNA (lambda digested) and 20 uL | <li>Mix 20 uL ssDNA (lambda digested) and 20 uL 4X SSC buffer</li> | ||
<li>Incubate in the dark at 70C for 5 minutes</li> | |||
<li>Incubate at 55C for 20 minutes</li> | |||
<li>Add 1 uL appropriate (10 uM) dye, A488-dc6 for sample 1 and Cy5-dc6 for sample 3</li> | <li>Add 1 uL appropriate (10 uM) dye, A488-dc6 for sample 1 and Cy5-dc6 for sample 3</li> | ||
<li>Incubate in the dark (RT) for 15 minutes</li> | <li>Incubate in the dark (RT) for 15 minutes</li> | ||
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</ol> | </ol> | ||
==Results== | |||
<gallery perrow=2 heights=250px widths=250px mode=packed-hover caption="Sample 2, Barcode 3 (Cy5-Red)"> | |||
File:20170712-emPCR8_Barcode3-Field4-SNAP_ch00.jpg|Field 4 | |||
File:20170712-emPCR8_Barcode3-Field5-SNAP_ch00.jpg|Field 5 | |||
File:20170712-emPCR8_Barcode3-Field6-SNAP_ch00.jpg|Field 6 | |||
</gallery> | |||
<gallery perrow=2 heights=250px widths=250px mode=packed-hover caption="Sample 3, Barcode 1 (A488-Cyan) and Barcode 3 (Cy5-Red)"> | |||
File:20170712-emPCR8_Barcode-v1v3-Field1-SNAP_ch00.jpg|Field 1 | |||
File:20170712-emPCR8_Barcode-v1v3-Field2-SNAP_ch00.jpg|Field 2 | |||
File:20170712-emPCR8_Barcode-v1v3-Field5-SNAP_ch00.jpg|Field 5 | |||
File:20170712-emPCR8_Barcode-v1v3-Field6-SNAP_ch00.jpg | |||
</gallery> | |||
[[Category:PosSeq]] [[Category:20170706]] | [[Category:PosSeq]] [[Category:20170706]] |
Revision as of 21:40, 12 July 2017
emPCR8 (Started Thursday July 6)
Protocol (From Yesterday)
- Dye hybridization
- Mix 20 uL ssDNA (lambda digested) and 20 uL 4X SSC buffer
- Incubate in the dark at 70C for 5 minutes
- Incubate at 55C for 20 minutes
- Add 1 uL appropriate (10 uM) dye, A488-dc6 for sample 1 and Cy5-dc6 for sample 3
- Incubate in the dark (RT) for 15 minutes
- Magnet pulldown, 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
- 20170712-emPCR8 Barcode3-Field4-SNAP ch00.jpg
Field 4
- 20170712-emPCR8 Barcode3-Field5-SNAP ch00.jpg
Field 5
- 20170712-emPCR8 Barcode3-Field6-SNAP ch00.jpg
Field 6
- 20170712-emPCR8 Barcode-v1v3-Field1-SNAP ch00.jpg
Field 1
- 20170712-emPCR8 Barcode-v1v3-Field2-SNAP ch00.jpg
Field 2
- 20170712-emPCR8 Barcode-v1v3-Field5-SNAP ch00.jpg
Field 5
- 20170712-emPCR8 Barcode-v1v3-Field6-SNAP ch00.jpg