03/16/11
- ✓ qRT-PCR: Pc-TF -/+ dox plate 3, 4
qRT-PCR
> 750 nM Primers (& cDNA dilution)
- NPPA 40A (1:10)
- CDKN2A 7C (1:10)
- MMP12 31B (1:10)
- THPO 33A (1:10)
> Plate 3: Templates, use 2 μL
- KAH126-1, 7/26/10
- KAH126-1 +dox, 7/26/10
- KAH128-8.3, 7/26/10
- KAH128-8.3 +dox, 7/26/10
- KAH129-4, 7/26/10
- KAH129-4 +dox, 7/26/10
- KAH132-8, 7/26/10
- KAH132-8 +dox, 7/26/10
> Plate 4: Templates, use 2 μL
- KAH154-2, 7/26/10
- KAH154-2 +dox, 7/26/10
- KAH156-5, 7/26/10
- KAH156-5 +dox, 7/26/10
- KAH157-1, 7/26/10
- KAH157-1 +dox, 7/26/10
- FTRx, 7/26/10
- FTRx +dox, 7/26/10
| Reagent |
1 rxn |
Primer mix (x25)
|
| diluted cDNA |
2.0 |
---
|
| SYBR Green mix |
7.5 |
187.5
|
| 750 nM primers |
3.0 |
75.0
|
| dH2O |
2.5 |
62.5
|
| |
15.0
|
--> Aliquot 39.0 primer mix into 1st well of each 3x set
--> Add 6.0 (2.0 x3) DNA to 39.0 primer mix
--> Aliquot 15.0 rxn mix to other 2 wells in each 3x set
Bio-Rad CFX96 qPCR (Kirschner lab)
--> Use Bio-Rad 96-well low profile plate MLL-9601 + Microseal "B" film
- 95°C/ 5 min.
- [95°C/ 15 sec, 58°C/ 15 sec, 72°C/ 15 sec] x45
- Melt curve range 58°C -> 95°C/ 0.5°C per step
|
Project name
Main project page
Previous entry Next entry
