Protocol 30007: End Point Analysis Assay - Kri1<em1(IMPC)J> Probe
Version 1.0

Notes

Mut = 122 bp

WT= 144 bp

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

Sequence

SEQ

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
33822 TGC ACC TAA AGA CTT TGT TTG G Mutant Forward A
33823 ATG AGA GGA AGG TCA TCT TGG A Wild type Forward A
33824 Fluorophore-1 TTG ATT TGT TTC TTT TGC ACA TC Quencher-1 MUT Probe
33825 Fluorophore-2 CCC GTC AGC TGC CAG TTC Quencher-2 WT Probe
33826 TTC CAG GAC AGC CAG AGC TA Common A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
33822 0.40 uM
33823 0.40 uM
33826 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.