Stock No: 028127
Protocol 22053: End Point Analysis Assay - Tbk1<em10(del2)Lutzy>
Version 2.0

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 = CA/--
Wild type = CA/CA

Sequence

ATGCAGAGCACCTCCAACCATCTGTGGCTCCTGTCTGATATCCTAGGCCAGGGGGCCACTG[ca]A
ATGTCTTCCGAGGAAGGCATAAGgtaggtccagtgctgcctgtcaggctcatctcgacctgg

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
24888 GAG CAC CTC CAA CCA TCT GT Forward A
24889 ACA GGC AGC ACT GGA CCT AC Reverse A
24893 Fluorophore-1 GGG CCA CTG CAA ATG TCT TC Quencher-1 WT Probe
24894 Fluorophore-2 GGG CCA CTG AAT GTC TTC C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
24888 0.40 uM
24889 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.