For in-depth product & services help, ask our
Technical Information Scientists
Stock No: 024531
Protocol 21996: End Point Analysis Assay - Lepre1<tm1.1Brle>
Version 3.0
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 = GCT/GCT
Heterozygote = CAT/GCT
Wild type = CAT/CAT
JAX Protocol
Protocol Primers
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| 18897 | GAA GGG CTG TGG GAT TCT CT | Forward | A | |||
| 18898 | AGG ATC CAG CGT GAA CCA C | Reverse | A | |||
| 18899 | Fluorophore-1 | GAG AAC CCA CAT GGA GTG AAG | Quencher-1 | WT Probe | ||
| 18900 | Fluorophore-2 | GAG AAC CCA GCT GGA GTG AAG | Quencher-2 | MUT Probe |
Reaction A
| Component | Final Concentration |
|---|---|
| Kapa Probe Fast QPCR | 1.00 X |
| ddH2O | |
| 18897 | 0.40 uM |
| 18898 | 0.40 uM |
| Wt Probe | 0.15 uM |
| Mutant Probe | 0.15 uM |
| DNA |
Cycling
| Step | Temp °C | Time | Note |
|---|---|---|---|
| 1 | 95.0 | -- | |
| 2 | 95.0 | -- | |
| 3 | 60.0 | -- | |
| 4 | -- | repeat steps 2-3 for 40 cycles | |
| 5 | 40.0 | -- | 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.