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 Pc-TF Genomics
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05/14/14
- RT-PCR: QC_plate, all cDNA batches
RT-PCR: QC_plate, all cDNA batches
- Summary: 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:
- Target gene wells: mCherry wells: 16x3 = 48; GAPD wells: 16x3 = 48
- cDNA dilution: 1:1000 for all reactions
- Plate layout: Plate_layouts_Carly.xlsx/QC_plate
cDNA Batches:
- K562_E001 (E = experimental, +PcTF)
- K562_C001 (C = control, no PcTF)
- K562_E002
- K562_C002
- K562_E003
- K562_C003
- SKNSH_E001
- SKNSH_C001
- SKNSH_E002
- SKNSH_C002
- SKNSH_E003
- SKNSH_C003
- U2OS_E001
- U2OS_C001
- U2OS_E002
- U2OS_C002
Target Gene Primer/Probe Master mixes (2 tubes)
| Reagent
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(Single well)
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Gene Target mCh
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Gene Target GAPD
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| 2x LC480 Probes Master |
(7.5 μL) |
360.0 |
360.0
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| 20 μM Forward primer |
(0.3 μL) |
14.4 |
14.4 GAPD primers
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| 20 μM Reverse primer |
(0.3 μL) |
14.4 |
0
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| 10 μM UPL probe |
(0.3 μL) |
14.4 |
14.4 GAPD probe
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| PCR H2O |
(0.1 μL) |
4.8 |
19.2
|
| Total vol. |
(8.5 μL) |
408.0 |
408.0
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Template Master Mixes (16 tubes)
| Reagent
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(Single well)
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cDNA Template
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no-template control
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| diluted Batch# cDNA |
(2.0 μL) |
12.0 |
0
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| PCR H2O |
(4.5 μL) |
27.0 |
0.0
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| Total vol. |
(6.5 μL) |
39.0 |
0.0
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Pc-TF Genomics
