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===AmpR13 and AmpR14 Results=== '''Labeling Efficiency''' {| 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="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" | AmpR13 | align="center" | 63.5 | align="center" | 3 | align="center" | 1.3 | align="center" | 3.8 | align="center" | 0.8 | align="center" | 1.6 | align="center" | 64 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | AmpR14 | align="center" | 70.8 | align="center" | 2.8 | align="center" | 1.4 | align="center" | 4.3 | align="center" | 0.7 | align="center" | 1.3 | align="center" | 77 |} '''Column Efficiency''' {| 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="45" | |style="font-weight:bold" width="65" | Pre ng/uL DNA |style="font-weight:bold" width="65" | Pre pmol/uL dye |style="font-weight:bold" width="65" | ng/uL DNA |style="font-weight:bold" width="65" | pmol/uL dye |style="font-weight:bold" width="71" | Column Yield (% DNA) |style="font-weight:bold" width="71" | Column Yield (% dye) |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | AmpR13 | align="center" | 72.9 | align="center" | 18.7 | align="center" | 63.5 | align="center" | 3 | align="center" | 75.7 | align="center" | 14.0 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | AmpR14 | align="center" | 68.9 | align="center" | 15 | align="center" | 70.8 | align="center" | 2.8 | align="center" | 89.4 | align="center" | 16.0 |} So it would appear that this trial worked. Since this is the best lead I've gotten, I'm going to go with it. It seems that VIGOROUS vortexing is needed, as well as much pipetting up and down. Note that for this trial I did not pipette the old dye mix (AmpR13) much before adding it to the DNA, but I mixed the DNA quite well for the fresher tube (AmpR14). This is another good sign that I'm on the right track, as the old tube used up the last of the dye and the new tube still has 3 uL left. I guess the next thing is to do a dye couple using the [[Daniel:Notebook/HiResChrPaint/2013-6-10#Nanodrop Results|V6S2 set]].
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