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==C Probe Binding Efficiency== '''Data Analysis Pipeline''' <ol> <li>Calculate average value of the samples for a given technical replicate</li> <li>Calculate Cycles above NTC using the NTC for that target (C1 or C2) on that specific run (Plates 4-6 or PosCon)</li> <li>Calculate the fold amplification over the 35 pg sample from PosCon (for C1 or C2)</li> <li>The amount in the wash was therefore estimated as 35 pg * the fold amplification (final result given in ng)</li> <li>Since 10 pmol C probe each was added, this is ~180 ng; each sample was calculated for percent in supernatant and the estimated percent bound (100-% in supernatant)</li> </ol> The table in the next section shows (some) of the intermediate values and final values. The graph is a scatter of the cycles above NTC and the % bound. For clarity, the "No C Probe" conditions are not shown (they are 0,0 and therefore outliers). [[Image:20170210-BeadsqPCR-CProbeBindingEfficency.png|600px]] ===C Probe Binding Results Table=== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#C4D79B;font-size:12pt;font-weight:bold" align="center" | width="300" height="45" | Name | width="65" | Average CT | width="90" | Abs(Difference) | width="68" | Cycles Above NTC | width="84" | Fold Amplification to 35 pg | width="65" | Est. Amt in Wash (ng) | width="90" | Estimated Fraction in Supernatant (%) | width="71" | Estimated Fraction Bound (%) |- style="font-size:12pt" | height="15" valign="bottom" | Room Temp-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.36 | align="center" align="center" valign="bottom" | 0.22 | align="center" align="center" valign="bottom" | 22.9 | align="center" align="center" valign="bottom" | 200.9 | align="center" align="center" valign="bottom" | 7.0 | align="center" align="center" valign="bottom" | 3.9 | align="center" align="center" valign="bottom" | 96.1 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | No Template-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.25 | align="center" align="center" valign="bottom" | 0.08 | align="center" align="center" valign="bottom" | 23.0 | align="center" align="center" valign="bottom" | 216.8 | align="center" align="center" valign="bottom" | 7.6 | align="center" align="center" valign="bottom" | 4.2 | align="center" align="center" valign="bottom" | 95.8 |- style="font-size:12pt" | height="15" valign="bottom" | 30C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.23 | align="center" align="center" valign="bottom" | 0.19 | align="center" align="center" valign="bottom" | 23.0 | align="center" align="center" valign="bottom" | 220.6 | align="center" align="center" valign="bottom" | 7.7 | align="center" align="center" valign="bottom" | 4.3 | align="center" align="center" valign="bottom" | 95.7 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | 40C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.60 | align="center" align="center" valign="bottom" | 0.3 | align="center" align="center" valign="bottom" | 22.7 | align="center" align="center" valign="bottom" | 170.1 | align="center" align="center" valign="bottom" | 6.0 | align="center" align="center" valign="bottom" | 3.3 | align="center" align="center" valign="bottom" | 96.7 |- style="font-size:12pt" | height="15" valign="bottom" | 50C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.15 | align="center" align="center" valign="bottom" | 0.12 | align="center" align="center" valign="bottom" | 23.1 | align="center" align="center" valign="bottom" | 232.3 | align="center" align="center" valign="bottom" | 8.1 | align="center" align="center" valign="bottom" | 4.5 | align="center" align="center" valign="bottom" | 95.5 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | 60C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 6.89 | align="center" align="center" valign="bottom" | 0.32 | align="center" align="center" valign="bottom" | 24.2 | align="center" align="center" valign="bottom" | 494.6 | align="center" align="center" valign="bottom" | 17.3 | align="center" align="center" valign="bottom" | 9.6 | align="center" align="center" valign="bottom" | 90.4 |- style="font-size:12pt" | height="15" valign="bottom" | 70C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.03 | align="center" align="center" valign="bottom" | 0.08 | align="center" align="center" valign="bottom" | 24.1 | align="center" align="center" valign="bottom" | 448.8 | align="center" align="center" valign="bottom" | 15.7 | align="center" align="center" valign="bottom" | 8.7 | align="center" align="center" valign="bottom" | 91.3 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | 80C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.00 | align="center" align="center" valign="bottom" | 0.03 | align="center" align="center" valign="bottom" | 24.1 | align="center" align="center" valign="bottom" | 459.8 | align="center" align="center" valign="bottom" | 16.1 | align="center" align="center" valign="bottom" | 8.9 | align="center" align="center" valign="bottom" | 91.1 |- style="font-size:12pt" | height="15" valign="bottom" | 90C Wash-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 7.16 | align="center" align="center" valign="bottom" | 0.07 | align="center" align="center" valign="bottom" | 23.9 | align="center" align="center" valign="bottom" | 411.6 | align="center" align="center" valign="bottom" | 14.4 | align="center" align="center" valign="bottom" | 8.0 | align="center" align="center" valign="bottom" | 92.0 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | No C Probe-C Probe fraction (C1) | align="center" align="center" valign="bottom" | 30.43 | align="center" align="center" valign="bottom" | 0.55 | align="center" align="center" valign="bottom" | 0.7 | align="center" align="center" valign="bottom" | 4.1 | align="center" align="center" valign="bottom" | 1.4 | align="center" align="center" valign="bottom" | 7.9 | align="center" align="center" valign="bottom" | 0.0 |- style="font-size:12pt" | height="15" valign="bottom" | Room Temp-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.34 | align="center" align="center" valign="bottom" | 0.09 | align="center" align="center" valign="bottom" | 23.3 | align="center" align="center" valign="bottom" | 164.3 | align="center" align="center" valign="bottom" | 5.7 | align="center" align="center" valign="bottom" | 3.2 | align="center" align="center" valign="bottom" | 96.8 |- style="background-color:#DA9694;font-size:12pt" | height="15" valign="bottom" | No Template-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.76 | align="center" align="center" valign="bottom" | 0.77 | align="center" align="center" valign="bottom" | 22.9 | align="center" align="center" valign="bottom" | 122.8 | align="center" align="center" valign="bottom" | 4.3 | align="center" align="center" valign="bottom" | 2.4 | align="center" align="center" valign="bottom" | 97.6 |- style="font-size:12pt" | height="15" valign="bottom" | 30C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.86 | align="center" align="center" valign="bottom" | 0.86 | align="center" align="center" valign="bottom" | 22.8 | align="center" align="center" valign="bottom" | 114.2 | align="center" align="center" valign="bottom" | 4.0 | align="center" align="center" valign="bottom" | 2.2 | align="center" align="center" valign="bottom" | 97.8 |- style="background-color:#DA9694;font-size:12pt" | height="15" valign="bottom" | 40C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.35 | align="center" align="center" valign="bottom" | 0.19 | align="center" align="center" valign="bottom" | 23.3 | align="center" align="center" valign="bottom" | 163.1 | align="center" align="center" valign="bottom" | 5.7 | align="center" align="center" valign="bottom" | 3.2 | align="center" align="center" valign="bottom" | 96.8 |- style="font-size:12pt" | height="15" valign="bottom" | 50C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.20 | align="center" align="center" valign="bottom" | 0.02 | align="center" align="center" valign="bottom" | 23.4 | align="center" align="center" valign="bottom" | 180.4 | align="center" align="center" valign="bottom" | 6.3 | align="center" align="center" valign="bottom" | 3.5 | align="center" align="center" valign="bottom" | 96.5 |- style="background-color:#DA9694;font-size:12pt" | height="15" valign="bottom" | 60C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.36 | align="center" align="center" valign="bottom" | 0.24 | align="center" align="center" valign="bottom" | 24.0 | align="center" align="center" valign="bottom" | 260.5 | align="center" align="center" valign="bottom" | 9.1 | align="center" align="center" valign="bottom" | 5.1 | align="center" align="center" valign="bottom" | 94.9 |- style="font-size:12pt" | height="15" valign="bottom" | 70C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.25 | align="center" align="center" valign="bottom" | 0.14 | align="center" align="center" valign="bottom" | 24.1 | align="center" align="center" valign="bottom" | 281.1 | align="center" align="center" valign="bottom" | 9.8 | align="center" align="center" valign="bottom" | 5.5 | align="center" align="center" valign="bottom" | 94.5 |- style="background-color:#DA9694;font-size:12pt" | height="15" valign="bottom" | 80C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.26 | align="center" align="center" valign="bottom" | 0.03 | align="center" align="center" valign="bottom" | 24.1 | align="center" align="center" valign="bottom" | 280.1 | align="center" align="center" valign="bottom" | 9.8 | align="center" align="center" valign="bottom" | 5.4 | align="center" align="center" valign="bottom" | 94.6 |- style="font-size:12pt" | height="15" valign="bottom" | 90C Wash-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 7.43 | align="center" align="center" valign="bottom" | 0.17 | align="center" align="center" valign="bottom" | 23.9 | align="center" align="center" valign="bottom" | 249.0 | align="center" align="center" valign="bottom" | 8.7 | align="center" align="center" valign="bottom" | 4.8 | align="center" align="center" valign="bottom" | 95.2 |- style="background-color:#DA9694;font-size:12pt" | height="15" valign="bottom" | No C Probe-C Probe fraction (C2) | align="center" align="center" valign="bottom" | 31.14 | align="center" align="center" valign="bottom" | 0.11 | align="center" align="center" valign="bottom" | 0.2 | align="center" align="center" valign="bottom" | 1.8 | align="center" align="center" valign="bottom" | 6.4 | align="center" align="center" valign="bottom" | 3.5 | align="center" align="center" valign="bottom" | 0.0 |} [[Category:ComboLock]] [[Category:20170209]]
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