PCR Conditions for Genotyping
You will receive primer sequences for the gene targeting mice. The following conditions are used for PCR genotyping for all our gene targeting mice. We'll email you if special conditions are used for your mice.
|
PCR Mix - Stocks |
One Reaction |
Ten Reactions (Make for 11 Reactions) |
| dNTP 10mM | 0.25 µl | 2.75 µl |
| 10x PCR Buffer | 1.25 µl | 13.75 µl |
| Primers 10 uM | 0.3 µl each | 3.3 µl each |
| Taq Polymerase | 0.06 µl | 0.66 µl |
| H2O | 9.48 µl | 104.28 µl |
| Total Volume | 12.02 µl | 124.74 µl |
| Add DNA: 0.5 µl |
|
PCR Cycles |
|
|
| 1. Denaturation: | 94°C | 3 minutes |
| 2. Denaturation: | 94°C | 30 seconds |
| 3. Annealing: | 55 °C | 30 seconds |
| 4. Extension: 2-4 steps X 35 cycles | 72 °C | 30 seconds |
| 5. Extension | 72°C | 5 minutes |