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==Indexing qPCR== *Perform one step qPCR using Indexing seq. primer {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> |- style="font-size:8pt" align="center" |style="background-color:#FFFF00;font-weight:bold;text-decoration:underline" width="126" height="11" | Content (50ul Capture) 4 rxn |style="background-color:#FFFF00;font-weight:bold" width="35" | Volume |style="background-color:#FFFF00;font-weight:bold" width="70" | Master Mix |style="background-color:#FFFF00;font-weight:bold" width="81" | ChIPK36me3 |style="background-color:#FFFF00;font-weight:bold" width="65" | Index |style="background-color:#FFFF00;font-weight:bold" width="75" | ChIPK27me3 |style="background-color:#FFFF00;font-weight:bold" width="65" | Index |style="text-decoration:none" align="center" width="65" | | align="center" width="65" | |- style="font-size:8pt" align="center" | height="11" | Captured DNA | align="center" | 27 | Do not add to MM | GM12878 | align="center" | 1 | GM12878 | align="center" | 10 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | 2X Kapa HiFi mix | align="center" | 50 | align="center" | 550 | H0 | align="center" | 2 | H0 | align="center" | 11 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | 10uM AmpF6.3Sol | align="center" | 4 | align="center" | 44 | H7 | align="center" | 3 | H7 | align="center" | 12 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | 10uM AmpR6.3Ind | align="center" | 4 | Do not add to MM | H14 | align="center" | 4 | H14 | align="center" | 13 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | H2O | align="center" | 15 | align="center" | 165 | H16 | align="center" | 9 | H16 | align="center" | 14 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | SYBR Green 50X | align="center" | 0.4 | align="center" | 4.4 | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | |- style="font-size:8pt" align="center" | align="center" height="11" | | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | | align="center" | |- style="background-color:#3366FF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="11" | Content (20ul Capture) 7rxn | Volume | Master Mix | WCE | Index | cDNA | Index | gDNA | Index |- style="font-size:8pt" align="center" |style="text-decoration:none" height="11" | Captured DNA | align="center" | 11 | Do not add to MM | GM12878 | align="center" | 15 | GM | align="center" | 22 | H7 | align="center" | 27 |- style="font-size:8pt" align="center" | height="11" | 2X Kapa SYBG qPCR master mix | align="center" | 50 | align="center" | 825 | P10 | align="center" | 16 | H0 | align="center" | 23 | H14 | align="center" | 28 |- style="font-size:8pt" align="center" | height="11" | 10uM AmpF6.3Sol | align="center" | 4 | align="center" | 66 | DF2 | align="center" | 17 | H7 | align="center" | 24 | H16 | align="center" | 29 |- style="font-size:8pt" align="center" | height="11" | 10uM AmpR6.3Ind | align="center" | 4 | Do not add to MM | H0 | align="center" | 18 | H14 | align="center" | 25 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | H2O | align="center" | 31 | align="center" | 511.5 | H7 | align="center" | 19 | H16 | align="center" | 26 | align="center" | | align="center" | |- style="font-size:8pt" align="center" | height="11" | SYBR Green 50X | align="center" | 0.4 | align="center" | 6.6 | H14 | align="center" | 20 | align="center" | | align="center" | | align="center" | | align="center" | |- style="font-size:8pt" align="center" | align="center" height="11" | | align="center" | | align="center" | | H16 | align="center" | 21 | align="center" | | align="center" | | align="center" | | align="center" | |} 95C 2min -> (98C 20S -> 58C 20S -> 72C 20S) x 6 -> (98C 10S -> 72C 20S) x40 Monitor the reactions and terminate the program right before the amplification curves reach the plateau.
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