Stock No: 030192
Protocol 21157: Probe Assay - Fdx1l<em1Murr>
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 = 13 bp deletion of ATCATGGCCGCCT

High G-C content

 

Sequence

CCCAGTGCATCACCTTTGACCTCACACGGCGCCCAAACGCTGCGTTTACTGTCGCTGAGGACGCGCAGAGCA
TGCTAGGTCTTCCGGTCACGTGGCCT[atcatggccgcct]CCATGGCCCGGGGCGTGAGCGCCCGGGTCCTGTTAC
GGGCGGCCGGGGGATCTTGGGGACCCAGGGCCGGGCACGCGGCGGTAACATCTAGGACGTTCGGGACTACAGG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
31858 CAG AGC ATG CTA GGT CTT CC Forward A
31859 CCT GGG TCC CCA AGA TCC Reverse A
31860 Fluorophore-1 CTA TCA TGG CCG CCT CC Quencher-1 WT Probe
31861 Fluorophore-2 CGT GGC CTC CAT GGC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
31858 0.40 uM
31859 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.