User:Karmella Haynes/Notebook/PcTF Genomics/2014/05/14

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05/14/14

  • RT-PCR: QC_plate, all cDNA batches



RT-PCR: QC_plate, all cDNA batches

  • Run mCh, GAPD reactions on all cDNA batches, all cell lines: 16 total
  • Will be used to compare cDNA quality, PcTF expression, and generate reference values
  • Experiment file name:
  • mCherry wells: 16x3 = 48
  • GAPD wells: 16x3 = 48
  • cDNA dilution: 1:1000 for all reactions


cDNA Batches:

  1. K562_E001
  2. K562_C001
  3. K562_E002
  4. K562_C002
  5. K562_E003
  6. K562_C003
  7. SKNSH_E001
  8. SKNSH_C001
  9. SKNSH_E002
  10. SKNSH_C002
  11. U2OS_E001
  12. U2OS_C001
  13. U2OS_E002
  14. U2OS_C002
  15. U2OS_E003
  16. U2OS_C003


Primer/Probe Master mixes (2 tubes)

Reagent (Single well) Gene Target mCh Gene Target GAPD (x10)
2x LC480 Probes Master(7.5 μL)360.0360.0
20 μM Forward primer(0.3 μL)14.414.4 GAPD primers
20 μM Reverse primer(0.3 μL)14.40
10 μM UPL probe(0.3 μL)14.414.4 GAPD probe
PCR H2O(0.1 μL)4.819.2
Total vol.(8.5 μL)408.0408.0


Template Master Mixes (16 tubes)

Reagent (Single well) cDNA Template no-template control
diluted Batch# cDNA(2.0 μL)12.00
PCR H2O(4.5 μL)27.00.0
Total vol.(6.5 μL)39.00.0




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