Stock No: 003612
Protocol 22065: End Point Analysis Assay - Trak1<hyrt>
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 = --------------------/--------------------
Heterozygote =cgtgcggagcagcgagagcc/--------------------
Wild type = cgtgcggagcagcgagagcc/cgtgcggagcagcgagagcc

Mutation is a 20bp deletion

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19050 CCC TCC TTC GAG TTC AAG TG Forward A
19051 AGG TCC CTG TTC CTC AGT CA Reverse A
19052 Fluorophore-1 TGC GGA GCA GCG AGA G Quencher-1 WT Probe
19053 Fluorophore-2 GAA AAG AGG CCA AAC AGA TGT GT Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
19050 0.40 uM
19051 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.