Stock No: 028435
Protocol 17690: End Point Analysis Assay - Tardbp<em#(G348C)Lutzy>-EP
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 = T/T
Heterozygote = G/T
Wild type = G/G

Sequence

CATTAACCCAGCGATGATGGCTGCGGCTCAGGCAGCGTTGCAGAGCAGTTGGGGTATGATGG
GCATGTTAGCCAGCCAGCAGAACCAGTCG(g/t)GCCCATCTGGGAATAACCAAAGCCAGGGCA
GCATGCAGAGGGAACCAAATCAGGCTTTTGGTTCTGGAAATAATTCCTACAGTGG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
25847 GAG CAG TTG GGG TAT GAT GG Forward A
25848 CAC CAG AAT TAG AAC CAC TGT AGG Reverse A
25849 Fluorophore-1 ACC AGT CGG GCC CAT CT Quencher-1 WT Probe A
25850 Fluorophore-2 AAC CAG TCG TGC CCA TCT G Quencher-2 MUT Probe A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
25847 0.40 uM
25848 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.