User:Karmella Haynes/Notebook/PcTF Genomics/2014/01/27

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01/27/14

  • qRT-PCR experiment - PlateK562_1/ U2OS

qRT-PCR - Plate K562_1/ U2OS


  Template Gene Target
Rxn 1:treated cellsKIT, primers 13/14
Rxn 2:treated cellsTNFRSF11A, primers 15/16
Rxn 3:treated cellsEGFR, primers 17/18
Rxn 4:treated cellsWT1, primers 19/20
Rxn 5:treated cellsHLF, primer 21/22
Rxn 6:treated cellsBCL6, primer 23/24
Rxn 7:treated cellsmCh
Rxn 8:treated cellsref. gene, GAPD
Rxn 9:untreated cellsKIT, primers 13/14
Rxn 10:untreated cellsTNFRSF11A, primers 15/16
Rxn 11:untreated cellsEGFR, primers 17/18
Rxn 12:untreated cellsWT1, primers 19/20
Rxn 13:untreated cellsHLF, primer 21/22
Rxn 14:untreated cellsBCL6, primer 23/24
Rxn 15:untreated cellsmCh
Rxn 16:untreated cellsref. gene, GAPD
Rxn 17:no templateKIT, primers 13/14
Rxn 18:no templateTNFRSF11A, primers 15/16
Rxn 19:no templateEGFR, primers 17/18
Rxn 20:no templateWT1, primers 19/20
Rxn 21:no templateHLF, primer 21/22
Rxn 22:no templateBCL6, primer 23/24
Rxn 23:no templatemCh
Rxn 24:no templateref. gene, GAPD


Primer/Probe Master Mixes (8 total)

Reagent (Single well) Gene Target 1 - 7 (x10) Gene Target GAPD (x10)
2x LC480 Probes Master(7.5 μL)7575
20 μM Forward primer(0.3 μL)33.0 GAPD primers*
20 μM Reverse primer(0.3 μL)3---
10 μM UPL probe(0.3 μL)33.0 GAPD UPL probe*
PCR H2O(0.1 μL)14
Total vol.(8.5 μL)8585



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