Stock No: 003507
Protocol 22018: End Point Analysis Assay - Npr3<lgj-2J>End Point
Version 3.0

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 bp
Heterozygote = C/T
Wild type = C/C

Sequence

GGTGAGGCCGGCCATCGAGTACGCGCTGCGTAGCGTGGAGGGCAATGGCACCGGGAGGAAG
CTGCTGCCGCCAGGCACTCGCTTC(c/t)AGGTGGCCTACGAAGACTCGGACTGCGGCAACCG
CGCGCTCTTCAGTCTTGTGGACCGCGTGGCGGCGGCGCGCGGCGCCAAGCCGG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18505 GTA GCG TGG AGG GCA ATG Forward A
18506 ACA AGA CTG AAG AGC GCG Reverse A
18507 Fluorophore-1 ACT CGC TTC CAG GTG GCC T Quencher-1 WT Probe
18508 Fluorophore-2 ACT CGC TTC TAG GTG GCC T Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
18505 0.40 uM
18506 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.