Stock No: 002379
Protocol 21939: End Point Analysis Assay - Dmd<mdx-5Cv>
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 = T
Wild type = A

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
17920 AGT TCA TTG ATG GAG ACG GAA G Forward A
17921 CAT GAG CAT GAA ACT GTT CTT TCA C Reverse A
17922 Fluorophore-1 CGA GCA CAA GGA GAG ATT TCA AA Quencher-1 WT Probe
17923 Fluorophore-2 CGA GCA CAA GGT GAG ATT TCA AAT Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
17920 0.40 uM
17921 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.