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Validation/LC Conversion data attached to rec 126211
Transgene = 348 bp
Internal Control = 324 bp
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| 13063 | GGC GTG TCA CTC ATC CTT TT | Transgene Forward | ||||
| 13064 | GGG ACA CCC CCT CCT AGA AT | Transgene Reverse | ||||
| oIMR7338 | CTA GGC CAC AGA ATT GAA AGA TCT | Internal Positive Control Forward | ||||
| oIMR7339 | GTA GGT GGA AAT TCT AGC ATC ATC C | Internal Positive Control Reverse |
| Component | Final Concentration |
|---|---|
| ddH2O | |
| Kapa 2G HS buffer | 1.00 |
| MgCl2 | 2.00 |
| dNTP KAPA | 0.20 |
| oIMR7338 | 0.25 |
| oIMR7339 | 0.25 |
| 13063 | 0.25 |
| 13064 | 0.25 |
| 10x Loading Dye | 0.50 |
| Kapa 2G HS taq polym | 0.01 |
| EvaGreen | 1.00 |
| DNA |
| Step | Temp °C | Time | Note |
|---|---|---|---|
| 1 | 94.0 | -- | |
| 2 | 94.0 | -- | |
| 3 | 65.0 | -- | -1.5 C per cycle decrease |
| 4 | 68.0 | -- | |
| 5 | -- | repeat steps 2-4 for 10 cycles | |
| 6 | 94.0 | -- | |
| 7 | 50.0 | -- | |
| 8 | 72.0 | -- | |
| 9 | -- | repeat steps 6-8 for 28 cycles | |
| 10 | 72.0 | -- | |
| 11 | 10.0 | -- | hold |
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