Stock No: 002343
Protocol 18078: End Point Analysis Assay - Otc<spf>
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 = A/A
Heterozygote = C/A
Wild type = C/C

This strain is x-linked

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18357 GGA GGA CAC CCT TCC TTT CT Forward A
18358 AGC GGT GTC TGT GAG ACT TTC Reverse A
18359 Fluorophore-1 CAC AAG ACA TTC ACT TGG GTG TG Quencher-1 Wild type A
18360 Fluorophore-2 CAC AAG ACA TTA ACT TGG GTG TG Quencher-2 MUT Probe A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
18357 0.40 uM
18358 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 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.