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)

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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
  1. Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)
    1. For AlexaFluor 488 label using 5 uL DMSO (stable for 1 month at 4C)
    2. VORTEX THESE SOLUTIONS VIGOROUSLY BEFORE USE EVERY TIME!
  2. Resuspend DNA in 20 uL labeling buffer (Component C)
  3. Denature DNA at 95C for 5 minutes and snap cool on ice; centrifuge briefly
  4. Add appropriate volume of dye to sample (see table 2); if necessary add labeling buffer to bring volume to 25 uL
  5. Incubate at 80C for 15 minutes; stop the reaction by plunging the tube into an ice bath
  6. Purify using a column (Centri-Sep recommended)
File:ULYSIS ReactionTable.png