Daniel:Notebook/HiResChrPaint/2013-7-9: Difference between revisions
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5. Incubate for 15 minutes at 80C, snap cool on ice | 5. Incubate for 15 minutes at 80C, snap cool on ice | ||
6. Purify using [[Daniel:Protocols/CentriSep|Centri-Sep Column]] | 6. Purify using [[Daniel:Protocols/CentriSep|Centri-Sep Column]] | ||
===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 20 uL | |||
|style="font-weight:bold" width="65" | pmol/uL DNA | |||
|style="font-weight:bold" width="65" | dye/probe | |||
|style="font-weight:bold" width="65" | dye/100 bp | |||
|style="font-weight:bold" width="65" | Base:Dye Ratio (:1) | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | AmpR4 | |||
| align="center" | 31.8 | |||
| align="center" | 1.4 | |||
| align="center" | 0.636 | |||
| align="center" | 1.9 | |||
| align="center" | 0.7 | |||
| align="center" | 1.45 | |||
| align="center" | 69 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | BLANK | |||
| align="center" | 66.6 | |||
| align="center" | 0.2 | |||
| align="center" | 1.332 | |||
| align="center" | 4.0 | |||
| align="center" | 0.0 | |||
| align="center" | 0.10 | |||
| align="center" | 1009 | |||
|} |
Revision as of 18:53, 9 July 2013
S2 Probe Prep (Started 5/29/2013)
ULYSIS Dye Coupling
Based on yesterday's results I think I'm going to see what happens if I use all 5 uL of dye available (see ULYSIS protocol) in one labeling. I'll use the oligo Noi gave me again, as I have a lot and it doesn't waste probe.
- Following ULYSIS protocol
1. Add 5 uL DMSO to Alexa dye 488 2. Add 1 uL DNA in 19 uL labeling buffer (Component C) 3. Denature DNA for 5 minutes at 95C; snap cool on ice 4. Add 5 uL dye to sample 5. Incubate for 15 minutes at 80C, snap cool on ice 6. Purify using Centri-Sep Column
Results
ng/uL DNA | pmol/uL dye | ug DNA in 20 uL | pmol/uL DNA | dye/probe | dye/100 bp | Base:Dye Ratio (:1) | |
AmpR4 | 31.8 | 1.4 | 0.636 | 1.9 | 0.7 | 1.45 | 69 |
BLANK | 66.6 | 0.2 | 1.332 | 4.0 | 0.0 | 0.10 | 1009 |