09/28/10
- ✓ RT-PCR: biological replicates for 4-day dox induction of Pc-ATF subset
RT-PCR
> Pc-ATF biological replicates (30 total)
--> Templates (6/15, 6/18, 6/23/10)
- KAH126-1 -dox
- KAH126-1 +dox
- KAH126-3 -dox
- KAH126-3 +dox
- KAH132-8 -dox
- KAH132-8 +dox
- KAH154-2 -dox
- KAH154-2 +dox
- FTRx -dox
- FTRx +dox
--> Primers
- p16INK 7C (94 bp), (0.5 μL 1:1 cDNA) (30 rxns)
- GAPDH 21A (100 bp), (0.5 μL 1:1000 cDNA) (30 rxns)
- MMP12 31A (127 bp), (5.0 μL 1:100 cDNA) (30 rxns)
- mCh (100 bp), (1.0 μL 1:1000 cDNA) (30 rxns)
- THPO 33A (103 bp)
- TNF 34A (119 bp)
Reagent |
p16, GAPDH |
mCh |
MMP12, THPO, TNF
|
cDNA |
0.5 |
1.0 |
5.0
|
10 μM primers |
1.0 |
1.0 |
1.0
|
2x GoTaq Green |
10.0 |
10.0 |
10.0
|
dH2O |
8.5 |
8.0 |
3.5
|
|
20.0 μL
|
--> PCR (96-well)
- 95°C/ 3 min.
- [95°C/ 30 sec., 57°C/ 30 sec., 72°C/ 30 sec.] x32
- 72°C/ 3 min.
- 4°C/ ∞
> Conclusions: Looks like MMP12 is induced just by the KAH126 proteins. Cool! Repeat MMP12 and p16 with 2x more template
|