Stock No: 028694
Protocol 19389: End Point Analysis Assay - Chchd10<em5(1ins)Lutzy>-EP-alt1
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 = A/A
Heterozygote = -/A
Wild type = -/-

Mutation is an A insertion

 

Sequence

Ccactggaccctggagtagaggggtttattcccggagcattaaccttgtttctctcctcttg
cttcctgcagccgccca[a]GCCCCGCCTCCTGCGCACCCACCACCCTCAGCGGCCGCCCCGGC
ACCTGCCGCTCCCGGCCAGCCGGGTCTTATGGCTC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
26910 Fluorophore-1 CGC CCA GCC CCG Quencher-1 WT Probe
26911 Fluorophore-2 CGC CCA AGC CCC G Quencher-2 MUT Probe
28586 CCG GAG CAT TAA CCT TGT TT Forward A
28587 TGG TGG GTG CGC AGG A Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
28586 0.40 uM
28587 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 -- repeat steps 2-3 for 40 cycles
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.