Stock No: 012810
Protocol 21956: End Point Analysis Assay - Enpp1<asj>-EP
Version 4.0

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 = A/A
Heterozygote = T/A
Wild type = T/T

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19913 CCT ATG TAC CCT ACC AAG ACG Forward A
19914 CTG AAG TTG AAT TAT GCC CTG G Reverse A
19915 Fluorophore-1 CAT TAC AGC ATC GTC ACA GTA AGT AGG Quencher-1 WT Probe
19916 Fluorophore-2 CAT TAC AGC ATC GAC ACA GTA AGT AGG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
19913 0.40 uM
19914 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.