Stock No: 023840
Protocol 22020: End Point Analysis Assay - Nubp1<m1Nisw>
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

Wt=C/C

Het=C/A

Hom=A/A

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18165 ACG CAG AAG CAA AAT CAA GC Forward A
18166 AAT CCT CAG ATC CTG GCA CA Reverse A
18167 Fluorophore-1 CCC TCC CAC GAC TGG GG Quencher-1 WT Probe
18168 Fluorophore-2 CCC TCC CAA GAC TGG GG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
18165 0.40 uM
18166 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.