Daniel:Notebook/PosSequencing/2017-5-22: Difference between revisions
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<li>Image on microscope</li> | <li>Image on microscope</li> | ||
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==Results== | |||
<gallery perrow=4 heights=300px widths=300px caption="Results-Sample 1 (A488) and Sample 3 (Cy5); A488-Green; Cy5-Red"> | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field1-SNAP_ch00.jpg|Sample 1 Field 1 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field2-SNAP_ch00.jpg|Sample 1 Field 2 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field3-SNAP_ch00.jpg|Sample 1 Field 3 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field4-Cy5-SNAP_ch00.jpg|Sample 1 Field 4-Cy5 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field1-SNAP_ch00.jpg|Sample 2 Field 1 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field2-SNAP_ch00.jpg|Sample 2 Field 2 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field3-SNAP_ch00.jpg|Sample 2 Field 3 | |||
File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field3-Cy5-SNAP_ch00.jpg|Sample 2 Field 3-A488 |
Revision as of 17:21, 22 May 2017
Single Stranded Probe Binding Test (Started Friday May 19)
Protocol (Continued from Saturday May 20)
- Dye hybridization
- Mix 20 uL ssDNA (lambda digested) and 20 uL dsDNA (original, non-digested template)
- 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
- Image on microscope
Results
<gallery perrow=4 heights=300px widths=300px caption="Results-Sample 1 (A488) and Sample 3 (Cy5); A488-Green; Cy5-Red"> File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field1-SNAP_ch00.jpg|Sample 1 Field 1 File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field2-SNAP_ch00.jpg|Sample 1 Field 2 File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field3-SNAP_ch00.jpg|Sample 1 Field 3 File:20170522-LambdaExonucleaseDigestion.lif_Sample1-Field4-Cy5-SNAP_ch00.jpg|Sample 1 Field 4-Cy5 File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field1-SNAP_ch00.jpg|Sample 2 Field 1 File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field2-SNAP_ch00.jpg|Sample 2 Field 2 File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field3-SNAP_ch00.jpg|Sample 2 Field 3 File:20170522-LambdaExonucleaseDigestion.lif_Sample2-Field3-Cy5-SNAP_ch00.jpg|Sample 2 Field 3-A488