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Stock No: 025250
Protocol 12037: End Point Analysis Assay - Tg(Csk*)2Weis
Version 1.0
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
Mutant =GG
Heterozygote =AC/GG
Wild type = AC
JAX Protocol
Protocol Primers
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| 20003 | GAA CTC CTG ACC ACC CTG TC | Forward | ||||
| 20004 | GCT CGC ATG ACT TGT TTA AAG TTG | Reverse | ||||
| 20005 | Fluorophore-1 | TCT ACA TCG TCA CAG AGT ACA TGG | Quencher-1 | WT Probe | ||
| 20006 | Fluorophore-2 | TCT ACA TCG TCG GAG AGT ACA TGG | Quencher-2 | MUT Probe |
Reaction A
| Component | Final Concentration |
|---|---|
| Kapa Probe Fast QPCR | 0.42 |
| ddH2O | |
| Primer 1 | 0.03 |
| Primer 2 | 0.03 |
| Primer 3 | 0.02 |
| Primer 4 | 0.02 |
| DNA |
Cycling
| Step | Temp °C | Time | Note |
|---|---|---|---|
| 1 | 95.0 | -- | |
| 2 | 92.0 | -- | |
| 3 | 60.0 | -- | |
| 4 | -- | repeat steps 2-3 for 40 cycles | |
| 5 | 4.0 | -- | Forever |
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.