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==Capturing Remaining Gimelbrant lab Sample== *Add 2 drops of mineral oil to prevent evaporation *Samples stored at tall -20C Alan Rack white box red label "Gimelbrant Lab" {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> |- style="background-color:#BFBFBF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" width="270" height="13" | ChIPK36me3 | width="121" | Concentration ng/ul | width="41" | Vol | width="83" | Vol of probes | width="121" | 10X Ampligase Buffer | width="39" | H2O | width="33" | Total |- style="font-size:8pt" align="center" |style="text-decoration:none" height="13" | GM12878 | align="center" | 0.90 | align="center" | 40.00 | align="center" | 0.45 | align="center" | 2 | align="center" | 7.6 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H0 | align="center" | 1.00 | align="center" | 40.00 | align="center" | 0.50 | align="center" | 2 | align="center" | 7.5 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H7 | align="center" | 0.60 | align="center" | 40.00 | align="center" | 0.30 | align="center" | 2 | align="center" | 7.7 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H14 | align="center" | 0.50 | align="center" | 40.00 | align="center" | 0.25 | align="center" | 2 | align="center" | 7.8 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H16 | align="center" | 0.60 | align="center" | 40.00 | align="center" | 0.30 | align="center" | 2 | align="center" | 7.7 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | | | | align="center" | | align="center" | | align="center" | | align="center" | |- style="background-color:#BFBFBF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="13" | ChIPK27me3 | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |- style="font-size:8pt" align="center" |style="text-decoration:none" height="13" | GM12878 | align="center" | 1.20 | align="center" | 40.00 | align="center" | 0.60 | align="center" | 2 | align="center" | 7.4 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H0 | align="center" | 2.4 | align="center" | 40.00 | align="center" | 1.20 | align="center" | 2 | align="center" | 6.8 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H7 | align="center" | 1.2 | align="center" | 40.00 | align="center" | 0.60 | align="center" | 2 | align="center" | 7.4 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H14 | align="center" | 0.5 | align="center" | 40.00 | align="center" | 0.25 | align="center" | 2 | align="center" | 7.8 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | H16 | align="center" | 1.3 | align="center" | 40.00 | align="center" | 0.65 | align="center" | 2 | align="center" | 7.4 | align="center" | 50 |- style="font-size:8pt" align="center" | height="13" | | | | align="center" | | align="center" | | align="center" | | align="center" | |- style="background-color:#BFBFBF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="13" | WCE | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |- style="font-size:8pt" align="center" |style="text-decoration:none" height="13" | GM12878 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | P10 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | DF2 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H0 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H7 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H14 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H16 | align="center" | 35.39 | align="center" | 5.65 | align="center" | 2.49 | align="center" | 2 | align="center" | 9.9 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | | | | align="center" | | align="center" | | align="center" | | align="center" | |- style="background-color:#BFBFBF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="13" | cDNA | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |- style="font-size:8pt" align="center" |style="text-decoration:none" height="13" | GM | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H0 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H7 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H14 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H16 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | | | | align="center" | | align="center" | | align="center" | | align="center" | |- style="background-color:#BFBFBF;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="13" | gDNA | Concentration ng/ul | Vol | Vol of probes | 10X Ampligase Buffer | H2O | Total |- style="font-size:8pt" align="center" |style="text-decoration:none" height="13" | H7 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H14 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |- style="font-size:8pt" align="center" | height="13" | H16 | align="center" | 38.12 | align="center" | 5.25 | align="center" | 2.49 | align="center" | 2 | align="center" | 10.3 | align="center" | 20 |} *When adding regents to the tube make sure you penetrate the oil layer Store under βCES36K 95C 2min -> -0.2C/sec to 60C -> 60C 24h -> add 1ul/2.5ul(for 50ul rxn) SLN mix (dNTP 5uM, Stoffel 2U/ul, AmpLigase 1U/ul) -> 60C 18h -> 94C 1min -> 37c 1min -> add 2ul(5ul for 50ul rxn) Exo I&III -> 37C 2h -> 90C 5min -> 4C hold ===Prepare SLN Mix=== *Dilute 1mM dNTP 1 in 100 to 10uM dNTP {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> |- style="font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" width="81" height="15" | SLN Mix for 20ul Rxn |style="text-decoration:none" width="112" | 1x | width="83" | 16x |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | dNTP (10uM) | align="center" | 0.5 | align="center" | 8 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | Stoffel (10U/ul) | align="center" | 0.2 | align="center" | 3.2 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | Ampligase (5U/ul) | align="center" | 0.2 | align="center" | 3.2 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | H2O | align="center" | 0.1 | align="center" | 1.6 |- style="font-size:8pt" align="center" | align="center" height="15" | | align="center" | | align="center" | |- style="font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="15" | SLN Mix for 50ul Rxn |style="text-decoration:none" | 1x | 11x |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | dNTP (10uM) | align="center" | 1.25 | align="center" | 13.75 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | Stoffel (10U/ul) | align="center" | 0.5 | align="center" | 5.5 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | Ampligase (5U/ul) | align="center" | 0.5 | align="center" | 5.5 |- style="font-size:8pt" align="center" |style="font-weight:bold" height="15" | H2O | align="center" | 0.25 | align="center" | 2.75 |} ===Prepare Exo I and Exo III Mix=== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext> |- style="background-color:#FFFF00;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" width="81" height="15" | Exo I & III for 20ul Rxn |style="text-decoration:none" width="112" | 1X | width="83" | 16x |- style="font-size:8pt" align="center" | height="15" | Exo I (20U/ul) | align="center" | 1 | align="center" | 16 |- style="font-size:8pt" align="center" | height="15" | ExoIII (200U/ul) | align="center" | 1 | align="center" | 16 |- style="font-size:8pt" align="center" | align="center" height="15" | | align="center" | | align="center" | |- style="background-color:#FFFF00;font-size:8pt;font-weight:bold" align="center" |style="text-decoration:underline" height="15" | Exo I & III for 50ul Rxn |style="text-decoration:none" | 1X | 11x |- style="font-size:8pt" align="center" | height="15" | Exo I (20U/ul) | align="center" | 2.5 | align="center" | 6.25 |- style="font-size:8pt" align="center" | height="15" | ExoIII (200U/ul) | align="center" | 2.5 | align="center" | 6.25 |}
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