Daniel:Notebook/HiResChrPaint/2013-7-11: Difference between revisions

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  4. Incubate for 15 minutes (AmpR9) or 20 minutes (AmpR10) at 80C, snap cool on ice
  4. Incubate for 15 minutes (AmpR9) or 20 minutes (AmpR10) at 80C, snap cool on ice
  5. Purify using [[Daniel:Protocols/CentriSep|Centri-Sep Column]]
  5. 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" | &nbsp;
|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="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="65" | Base:Dye Ratio (:1)
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | AmpR9
| align="center" | 64.6
| align="center" | 0.6
| align="center" | 1.3
| align="center" | 3.9
| align="center" | 0.2
| align="center" | 0.3
| align="center" | 326
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | AmpR10
| align="center" | 71.1
| align="center" | 0.4
| align="center" | 1.4
| align="center" | 4.3
| align="center" | 0.1
| align="center" | 0.2
| align="center" | 539
|}
It would seem that I can always tell beforehand whether a yield will be good or bad, by the color of
the solution post incubation.  If the solution has a faint green color it will be a good yield.  If
the solution remains clear it will be a bad yield.  Therefore I'm going to ignore columns for future
trials (yes, it looks like I need more troubleshooting).  That will save on time and $$, and hopefully
I'll get to the bottom of this before long.

Revision as of 19:46, 11 July 2013

S2 Probe Prep (Started 5/29/2013)

Back to Calendar

ULYSIS Dye Coupling (Test 6)

Should be the last test.  I'm going to test at 15 minutes and 20 minutes incubation time, 1 uL each.  If the
results go well this morning I will be labeling the V6S2 set this afternoon. 
1. Add 1 uL DNA in 19 uL labeling buffer (Component C)
2. Denature DNA for 5 minutes at 95C; snap cool on ice
3. Add 1 uL dye to sample; add 4 uL labeling buffer
4. Incubate for 15 minutes (AmpR9) or 20 minutes (AmpR10) at 80C, snap cool on ice
5. 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)
AmpR9 64.6 0.6 1.3 3.9 0.2 0.3 326
AmpR10 71.1 0.4 1.4 4.3 0.1 0.2 539
It would seem that I can always tell beforehand whether a yield will be good or bad, by the color of
the solution post incubation.  If the solution has a faint green color it will be a good yield.  If 
the solution remains clear it will be a bad yield.  Therefore I'm going to ignore columns for future 
trials (yes, it looks like I need more troubleshooting).  That will save on time and $$, and hopefully
I'll get to the bottom of this before long.