qPCR of Hirt and Puregene Extracted Dinn-1 Cells To Compare DiNV DNA, PIF3 and TPI Primers
Using PIF3 (virus) and TPI primers for this qPCR, using samples that were extracted here, puregene samples were added in with last cell DNA extraction
- Samples were kept on ice
- Reagents were thawed and kept on ice
- Master mixes for each primer were made and kept on ice until use
- TPI master mix
- 130ul Sso
- 13ul TPI F primer
- 13ul TPI R primer
- 78ul molecular grade water
- PIF3 master mix
- 130ul Sso
- 13ul PIF3 F primer
- 13ul PIF3 R primer
- 78ul molecular grade water
- 9ul of primer was added to each well of a qPCR plate
- 1ul of diluted (or not) sample was added to the appropriate well in the plate
- The wells were pipette mixed 10X with a multichannel to mix
- The plate was sealed with the biorad seal
- The plate was spun down for 3 minutes at 4000rpm
- The plate was brought to the qPCR machine and run on the p47 program
Plate layout here
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| A | 32 | 32 | 32 | 33 | 33 | 33 | 34 | 34 | 34 | 35 | 35 | 35 | TPI |
| B | 36 | 36 | 36 | 37 | 37 | 37 | 38 | 38 | 38 | 39 | 39 | 39 | |
| C | 32 | 32 | 32 | 33 | 33 | 33 | 34 | 34 | 34 | 35 | 35 | 35 | PIF 3 |
| D | 36 | 36 | 36 | 37 | 37 | 37 | 38 | 38 | 38 | 39 | 39 | 39 | |
| E | |||||||||||||
| F | |||||||||||||
| G | |||||||||||||
| H |
Sample info here
| well | sample | replicate | primer | type |
|---|---|---|---|---|
| A01 | 32 | 1 | TPI | Puregene |
| A02 | 32 | 2 | TPI | Puregene |
| A03 | 32 | 3 | TPI | Puregene |
| A04 | 33 | 1 | TPI | Puregene |
| A05 | 33 | 2 | TPI | Puregene |
| A06 | 33 | 3 | TPI | Puregene |
| A07 | 34 | 1 | TPI | Puregene |
| A08 | 34 | 2 | TPI | Puregene |
| A09 | 34 | 3 | TPI | Puregene |
| A10 | 35 | 1 | TPI | Puregene |
| A11 | 35 | 2 | TPI | Puregene |
| A12 | 35 | 3 | TPI | Puregene |
| B01 | 36 | 1 | TPI | hirt |
| B02 | 36 | 2 | TPI | hirt |
| B03 | 36 | 3 | TPI | hirt |
| B04 | 37 | 1 | TPI | hirt |
| B05 | 37 | 2 | TPI | hirt |
| B06 | 37 | 3 | TPI | hirt |
| B07 | 38 | 1 | TPI | hirt |
| B08 | 38 | 2 | TPI | hirt |
| B09 | 38 | 3 | TPI | hirt |
| B10 | 39 | 1 | TPI | hirt |
| B11 | 39 | 2 | TPI | hirt |
| B12 | 39 | 3 | TPI | hirt |
| C01 | 32 | 1 | PIF 3 | Puregene |
| C02 | 32 | 2 | PIF 3 | Puregene |
| C03 | 32 | 3 | PIF 3 | Puregene |
| C04 | 33 | 1 | PIF 3 | Puregene |
| C05 | 33 | 2 | PIF 3 | Puregene |
| C06 | 33 | 3 | PIF 3 | Puregene |
| C07 | 34 | 1 | PIF 3 | Puregene |
| C08 | 34 | 2 | PIF 3 | Puregene |
| C09 | 34 | 3 | PIF 3 | Puregene |
| C10 | 35 | 1 | PIF 3 | Puregene |
| C11 | 35 | 2 | PIF 3 | Puregene |
| C12 | 35 | 3 | PIF 3 | Puregene |
| D01 | 36 | 1 | PIF 3 | hirt |
| D02 | 36 | 2 | PIF 3 | hirt |
| D03 | 36 | 3 | PIF 3 | hirt |
| D04 | 37 | 1 | PIF 3 | hirt |
| D05 | 37 | 2 | PIF 3 | hirt |
| D06 | 37 | 3 | PIF 3 | hirt |
| D07 | 38 | 1 | PIF 3 | hirt |
| D08 | 38 | 2 | PIF 3 | hirt |
| D09 | 38 | 3 | PIF 3 | hirt |
| D10 | 39 | 1 | PIF 3 | hirt |
| D11 | 39 | 2 | PIF 3 | hirt |
| D12 | 39 | 3 | PIF 3 | hirt |