Stock No: 029033
Protocol 19791: End Point Analysis Assay - Chchd10<em11(del10)Lutzy>
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 = ----------/----------
Heterozygote = CAGCCCCGCC/----------
Wild type =  CAGCCCCGCC/CAGCCCCGCC

Sequence

CCCGGAGCATTAACCTTGTTTCTCTCCTCTTGCTTCCTGCAGCCGCC[cagccccgcc]TCCTGCGCACCCACCACCCTCAGC
GGCCGCCCCGGCACCTGCCGCTCCCGGCCAGCCGGGTCTTATGGCTCAGATGGCATCC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
26901 CCC GGA GCA TTA ACC TTG T Forward A
26902 GGA TGC CAT CTG AGC CAT AAG Reverse A
29254 Fluorophore-1 CCC AGC CCC GCC TC Quencher-1 WT Probe
29255 Fluorophore-2 AGC CGC CTC CTG CG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
26901 0.40 uM
26902 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.