Stock No: 023899
Protocol 11255: Standard PCR Assay - Insm1<tm1.1Mgn>
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 = ~550 bp
Heterozygote = 447 bp and ~550 bp
Wild type = 447 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18544 CCT TGT ACA ACC GAC AGC TCT Forward
18545 GTG CCC TGT ATC TGC TGT GC Reverse

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 94.0 --
2 94.0 --
3 65.0 -- -0.5 C per cycle decrease
4 68.0 --
5 -- repeat steps 2-4 for 10 cycles
6 94.0 --
7 60.0 --
8 72.0 --
9 -- repeat steps 6-8 for 28 cycles
10 72.0 --
11 10.0 -- hold
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.