Stock No: 035235
Protocol 39468: End Point Analysis Assay - Pex10<em1Lutzy>
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

>chr4:155070675+155070771 97bp CCACTCTGCACCCTATGCT TGTTGCACCACTCGGTGA

Mutant = C, T, G

Wild type = T, C, C

Sequence

CGTGTGCAGAACCCCACTCTGCACCCTATGCTTAGAGGAACGAAGACACTCCACGGCCACACCTTG(t/c)GG(c/t)(c/g)ATCTCTTCTGCTGGGAGTGCATCACCGAGTGGTGCAACACCAAGgtactcgcgctcccgggcctg

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
53879 CCA CTC TGC ACC CTA TGC T Forward A
53880 TGT TGC ACC ACT CGG TGA Reverse A
53882 Fluorophore-1 ACC TTG TGG CCA TCT CTT CTG Quencher-1 WT Probe
53883 Fluorophore-2 ACC TTG CGG TGA TCT CTT CTG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
53879 0.40 uM
53880 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.