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Notes
The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX).
To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility.
Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.
Expected Results
JAX Protocol
Protocol Primers
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| oIMR1436 | GGT AAA GAC CCA TGT GAC CC | Common | ||||
| oIMR1437 | TCC ACC TAG CCT GCC TGT AC | Mutant | ||||
| oIMR1438 | GGC TTG CCA CAT GAC AA | Wild type |
Reaction A
| Component | Final Concentration |
|---|---|
| ddH2O | |
| High Resolution Melt | 1.00 X |
| MgCl2 | 2.00 mM |
| oIMR1436 | 1.00 uM |
| oIMR1437 | 1.00 uM |
| oIMR1438 | 1.00 uM |
| DNA |
Cycling
| Step | Temp °C | Time | Note |
|---|---|---|---|
| 1 | 94.0 | -- | |
| 2 | 94.0 | -- | |
| 3 | 59.0 | -- | |
| 4 | 72.0 | -- | repeat steps 2-4 for 35 cycles |
| 5 | 72.0 | -- | |
| 6 | 10.0 | -- | hold |
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.
Strains Using This Protocol
This is the only strain that uses this protocol.