|
05/20/14
- qRT-PCR: Run "K562_1" plate for SKNSH and U2OS
RT-PCR: QC_plate, all cDNA batches
- System: Roche LC480 (machine & reagents)
- Summary: Complementary experiment; Look at expression of genes identified as unique targets for K562 cells
- Experiment file name: DualColorProbe_UPL_Haynes052014
- Wells per Target Gene: 4 cDNAs x 3 reps = 12
- Wells per cDNA Template: 6 genes x 3 reps = 18
- cDNA dilution: 1:10 for all reactions
- Plate layout: Plate_layouts_Carly.xlsx/K562_1
Target Genes & Oligos:
- MAPK14 - UPL-65, 1, 2 (20 μM separate primers)
- MAPK9 - UPL-57, 3, 4
- MAPK10 - UPL-1, 5, 6
- RET - UPL-3, 7, 8
- FGFR4 - UPL-1, 9, 10
- CDKN1A - UPL-1, 11, 12
Primer/Probe Master mixes (6 tubes)
| Reagent
|
(Single well)
|
Target Gene (x12.5)
|
Roche GAPD (x0)
|
| 2x LC480 Probes Master |
(7.5 μL) |
93.75 |
0.00
|
| 20 μM Forward primer |
(0.3 μL) |
3.75 |
0.00
|
| 20 μM Reverse primer |
(0.3 μL) |
3.75 |
0.00
|
| 10 μM UPL probe |
(0.3 μL) |
3.75 |
0.00
|
| PCR H2O |
(0.1 μL) |
1.25 |
0.00
|
| Total vol. |
(8.5 μL) |
106.25 |
0.00
|
cDNA Batches:
- SKNSH_E002 (E = experimental, +PcTF)
- SKNSH_C002 (C = control, no PcTF)
- U2OS_E002
- U2OS_C002
Template master mixes (4 tubes)
| Reagent
|
(Single well)
|
cDNA Template (x18.5)
|
no-template control (x0)
|
| diluted Batch# cDNA |
(2.0 μL) |
37.0 |
0.00
|
| PCR H2O |
(4.5 μL) |
83.3 |
0.00
|
| Total vol. |
(6.5 μL) |
120.3 |
0.00
|
|
Pc-TF Genomics
Main project page
Previous entry Next entry
