Daniel:Notebook/HiResChrPaint/2014-3-6: Difference between revisions
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>Djacobse |
>Djacobse |
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==Dye Labeling== | ==Dye Labeling== | ||
Samples: | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" valign="bottom" | |||
| width="65" height="15" | Sample | |||
| width="65" | Amount | |||
| width="65" | Dye | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| height="15" | V6S2(A+B) | |||
| 6.3 ug | |||
| align="center" | 488 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| height="15" | V6S3(A+B) | |||
| 6.9 ug | |||
| align="center" | 546 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| height="15" | Cot1 | |||
| 6 uL=6ug | |||
| align="center" | 488 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| height="15" | BSPP1B | |||
| 2.7 ug | |||
| align="center" | 546 | |||
|} | |||
*Following [[Daniel:Protocols/AlexaDyeCouple#ULYSIS|ULYSIS protocol]] | *Following [[Daniel:Protocols/AlexaDyeCouple#ULYSIS|ULYSIS protocol]] |
Revision as of 17:33, 6 March 2014
Overload FISH (Started 3/5/2014)
Dye Labeling
Samples:
Sample | Amount | Dye |
V6S2(A+B) | 6.3 ug | 488 |
V6S3(A+B) | 6.9 ug | 546 |
Cot1 | 6 uL=6ug | 488 |
BSPP1B | 2.7 ug | 546 |
- Following ULYSIS protocol
- Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)
- For AlexaFluor 488 label using 5 uL DMSO (stable for 1 month at 4C)
- VORTEX THESE SOLUTIONS VIGOROUSLY BEFORE USE EVERY TIME!
- Resuspend DNA in 20 uL labeling buffer (Component C)
- Denature DNA at 95C for 5 minutes and snap cool on ice; centrifuge briefly
- Add appropriate volume of dye to sample (see table 2); if necessary add labeling buffer to bring volume to 25 uL
- Incubate at 80C for 15 minutes; stop the reaction by plunging the tube into an ice bath
- Purify using a column (Centri-Sep recommended)
File:ULYSIS ReactionTable.png