Daniel:Notebook/HiResChrPaint/2013-7-25: Difference between revisions
Jump to navigation
Jump to search
>Djacobse (Created page with "=S2 Probe Prep (Started 7/23/2013)= Back to Main ==Dye Coupling== *Following [[D...") |
>Djacobse No edit summary |
||
Line 23: | Line 23: | ||
===Labeling Results=== | ===Labeling Results=== | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" | |||
| align="center" width="65" height="30" | | |||
|style="font-weight:bold" width="65" | ng/uL DNA | |||
|style="font-weight:bold" width="65" | pmol/uL dye | |||
|style="font-weight:bold" width="65" | ug DNA in 15 uL | |||
|style="font-weight:bold" width="71" | pmol/uL DNA | |||
|style="font-weight:bold" width="71" | dye/probe | |||
|style="font-weight:bold" width="71" | dye/100 bp | |||
|style="font-weight:bold" width="71" | Base:Dye Ratio (:1) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | V4S3 | |||
| align="center" | 106.6 | |||
| align="center" | 3.8 | |||
| align="center" | 1.6 | |||
| align="center" | 4.0 | |||
| align="center" | 1.0 | |||
| align="center" | 1.2 | |||
| align="center" | 85 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | V6S3 | |||
| align="center" | 133.8 | |||
| align="center" | 5.5 | |||
| align="center" | 2.0 | |||
| align="center" | 5.0 | |||
| align="center" | 1.1 | |||
| align="center" | 1.4 | |||
| align="center" | 74 | |||
|} | |||
Again, the labeling results weren't fantastic, but they should serve as workable probes for FISH experiments. |
Revision as of 19:14, 25 July 2013
S2 Probe Prep (Started 7/23/2013)
Dye Coupling
- Following ARES protocol
0. Dry samples in a vaccuum centrifuge
For each
1. Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature 2. Resuspend DNA in 5 uL nfH20 3. Add 5 uL DNA and 3 uL of sodium bicarbonate buffer in PCR tube 4. Denature samples for 5 minutes at 95C, then snap cool using ice box 5. Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds *dye cannot be saved for later use. Use immediately! 6. Add 2 uL dye to sample 7. Incubate in the dark for 1 hour 8. Add 10 uL nfH20 to sample 9. Purify using Centri Sep column 10. Measure concentrations using Microarray setting in nanodrop
Labeling Results
ng/uL DNA | pmol/uL dye | ug DNA in 15 uL | pmol/uL DNA | dye/probe | dye/100 bp | Base:Dye Ratio (:1) | |
V4S3 | 106.6 | 3.8 | 1.6 | 4.0 | 1.0 | 1.2 | 85 |
V6S3 | 133.8 | 5.5 | 2.0 | 5.0 | 1.1 | 1.4 | 74 |
Again, the labeling results weren't fantastic, but they should serve as workable probes for FISH experiments.