>Alan6017518 |
>Alan6017518 |
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| Monitor the reactions and terminate the program right before the amplification curves reach the plateau. | | Monitor the reactions and terminate the program right before the amplification curves reach the plateau. |
| ==Indexing PCR for 3 samples== | | ==Indexing PCR for 3 samples== |
| ==Indexing PCR with Kapa SYBR Fast==
| |
| {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext>
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| |- style="font-size:8pt" align="center"
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| |style="background-color:#FFFF00;font-weight:bold;text-decoration:underline" width="126" height="15" | Content (50ul Capture) 4 rxn
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| |style="background-color:#FFFF00;font-weight:bold" width="35" | Volume
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| |style="background-color:#FFFF00;font-weight:bold" width="70" | Master Mix
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| |style="background-color:#FFFF00;font-weight:bold" width="56" | ChIPK36me3
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| |style="background-color:#FFFF00;font-weight:bold" width="36" | Index
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| |style="background-color:#FFFF00;font-weight:bold" width="65" | ChIPK27me3
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| |style="background-color:#FFFF00;font-weight:bold" width="28" | Index
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| |style="text-decoration:none" align="center" width="65" |
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| | align="center" width="65" |
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|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | Captured DNA
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| | align="center" | 27
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| | Do not add to MM
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| | GM12878
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| | align="center" | 1
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| | GM12878
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| | align="center" | 10
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| | align="center" |
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| | align="center" |
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|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | 2X Kapa HiFi mix
| |
| | align="center" | 50
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| | align="center" | 495
| |
| |style="color:#FF0000" | H0
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| | align="center" | 2
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| | H0
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| | align="center" | 11
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| | align="center" |
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| | align="center" |
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|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | 10uM AmpF6.3Sol
| |
| | align="center" | 4
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| | align="center" | 39.6
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| | H7
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| | align="center" | 3
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| | H7
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| | align="center" | 12
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| | align="center" |
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| | align="center" |
| |
|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | 10uM AmpR6.3Ind
| |
| | align="center" | 4
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| | Do not add to MM
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| | H14
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| | align="center" | 4
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| | H14
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| | align="center" | 13
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| | align="center" |
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| | align="center" |
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|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | H2O
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| | align="center" | 15
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| | align="center" | 148.5
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| | H16
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| | align="center" | 9
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| | H16
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| | align="center" | 14
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| | align="center" |
| |
| | align="center" |
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| | height="15" | SYBR Green 50X
| |
| | align="center" | 0.4
| |
| | align="center" | 3.96
| |
| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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|
| |
| |- style="font-size:8pt" align="center"
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| | align="center" height="15" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
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| | align="center" |
| |
|
| |
| |- style="background-color:#3366FF;font-size:8pt;font-weight:bold" align="center"
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| |style="text-decoration:underline" height="15" | Content (20ul Capture) 7rxn
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| | Volume
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| | Master Mix
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| | WCE
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| | Index
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| | cDNA
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| | Index
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| | gDNA
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| | Index
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| |style="text-decoration:none" height="15" | Captured DNA
| |
| | align="center" | 11
| |
| | Do not add to MM
| |
| | GM12878
| |
| | align="center" | 15
| |
| |style="color:#FF0000" | GM
| |
| | align="center" | 22
| |
| | H7
| |
| | align="center" | 27
| |
|
| |
| |- style="font-size:8pt" align="center"
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| | height="15" | 2X Kapa SYBG qPCR master mix
| |
| | align="center" | 50
| |
| | align="center" | 715
| |
| | P10
| |
| | align="center" | 16
| |
| | H0
| |
| | align="center" | 23
| |
| | H14
| |
| | align="center" | 28
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| | height="15" | 10uM AmpF6.3Sol
| |
| | align="center" | 4
| |
| | align="center" | 57.2
| |
| |style="color:#FF0000" | DF2
| |
| | align="center" | 17
| |
| | H7
| |
| | align="center" | 24
| |
| | H16
| |
| | align="center" | 29
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| | height="15" | 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="15" | H2O
| |
| | align="center" | 31
| |
| | align="center" | 443.3
| |
| | H7
| |
| | align="center" | 19
| |
| | H16
| |
| | align="center" | 26
| |
| | align="center" |
| |
| | align="center" |
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| | height="15" | SYBR Green 50X
| |
| | align="center" | 0.4
| |
| | align="center" | 5.72
| |
| | H14
| |
| | align="center" | 20
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| | align="center" height="15" |
| |
| | align="center" |
| |
| | align="center" |
| |
| | H16
| |
| | align="center" | 21
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
|
| |
| |- style="font-size:12pt" valign="bottom"
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| | align="center" height="15" |
| |
| | align="center" |
| |
| | align="center" |
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| | align="center" |
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| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
| | align="center" |
| |
|
| |
| |- style="font-size:8pt" align="center"
| |
| |style="background-color:#CCFFCC;font-weight:bold;text-decoration:underline" height="15" | Content (15ul Capture) 3rxn
| |
| |style="background-color:#CCFFCC;font-weight:bold" | Volume
| |
| |style="background-color:#CCFFCC;font-weight:bold" | Master Mix
| |
| |style="background-color:#CCFFCC" | Type
| |
| |style="background-color:#CCFFCC;font-weight:bold" | Sample
| |
| |style="background-color:#CCFFCC;font-weight:bold" | Index
| |
| |style="font-weight:bold" align="center" |
| |
| |style="font-weight:bold" align="center" |
| |
| |style="font-weight:bold" align="center" |
| |
|
| |
| |-
| |
| |style="font-size:8pt;text-decoration:none" height="15" align="center" | Captured DNA
| |
| |style="font-size:8pt" align="center" align="center" | 15
| |
| |style="font-size:8pt" align="center" | Do not add to MM
| |
| |style="background-color:#FFFF00;font-size:8pt;font-weight:bold;text-decoration:underline" align="center" | ChIPK36me3
| |
| |style="font-size:8pt;text-decoration:none;color:#FF0000" align="center" | H0
| |
| |style="font-size:8pt" align="center" align="center" | 2
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
|
| |
| |-
| |
| |style="font-size:8pt" height="15" align="center" | 2X Kapa SYBG qPCR master mix
| |
| |style="font-size:8pt" align="center" align="center" | 50
| |
| |style="font-size:8pt" align="center" align="center" | 165
| |
| |style="background-color:#3366FF;font-size:8pt;font-weight:bold;text-decoration:underline" align="center" | WCE
| |
| |style="font-size:8pt;text-decoration:none;color:#FF0000" align="center" | DF2
| |
| |style="font-size:8pt" align="center" align="center" | 17
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
|
| |
| |-
| |
| |style="font-size:8pt" height="15" align="center" | 10uM AmpF6.3Sol
| |
| |style="font-size:8pt" align="center" align="center" | 4
| |
| |style="font-size:8pt" align="center" align="center" | 13.2
| |
| |style="background-color:#3366FF;font-size:8pt;font-weight:bold;text-decoration:underline" align="center" | cDNA
| |
| |style="font-size:8pt;text-decoration:none;color:#FF0000" align="center" | GM
| |
| |style="font-size:8pt" align="center" align="center" | 22
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
|
| |
| |-
| |
| |style="font-size:8pt" height="15" align="center" | 10uM AmpR6.3Ind
| |
| |style="font-size:8pt" align="center" align="center" | 4
| |
| |style="font-size:8pt" align="center" | Do not add to MM
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
|
| |
| |-
| |
| |style="font-size:8pt" height="15" align="center" | H2O
| |
| |style="font-size:8pt" align="center" align="center" | 27
| |
| |style="font-size:8pt" align="center" align="center" | 89.1
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
|
| |
| |-
| |
| |style="font-size:8pt" height="15" align="center" | SYBR Green 50X
| |
| |style="font-size:8pt" align="center" align="center" | 0.4
| |
| |style="font-size:8pt" align="center" align="center" | 1.32
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
| |
| |style="font-size:12pt" align="center" valign="bottom" |
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|
| |
| |}
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| *The 3 samples highlighted in red will use 15ul from the 1st round PCR (purified product) as the 2nd round PCR is already amplified with Kapa HiFi.
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| *First add Index primer
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