Protocol 45857: Probe Assay - Zfp524<em1(IMPC)J>
Version 1.0


Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.


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=  92 bp

Wild Type = 92 bp



Large deletion:  Mutant sequence with junction in uppercase



This mutation is a 1202 bp deletion beginning at Chromosome 7 position 5,017,385 bp and ending after 5,018,586 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
66632 CCT CCT GTG GAC TGA GAA GC Wild type Forward A
66633 CAG GCA CCA TGA CTG AAC AC Wild type Reverse A
66634 Fluorophore-1 ATC CCT GCC TAC TGA CTT CCA Quencher-1 WT Probe
66635 TTA GAA GGA ATA GGT GAG ATA CTG G Mutant Forward A
66636 AGA TTG GGC AAA GGC TTG TT Mutant Reverse A
66637 Fluorophore-2 CCC TAG CTT GGT GCC AAA GAA CC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
66632 0.40 uM
66633 0.40 uM
66635 0.40 uM
66636 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM


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.