Stock No: 003556
Protocol 40843: Probe Assay - Mx1-cre Probe
Version 2.0

Notes

This assay cannot distinguish hemi from hom

 

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

Tg=  124 bp

IPC = 132 bp

Sequence

Tg Sequence: ATATGTGGTGGGGGTGTGGGCACAGCAAGCTCAGGCTTTTTCACGGTTTCAATTCTCCTCTGGAAGGGGCGGAGCCAGCACTATTTAAAGAGGTCTGAGCCCTGGCAGggggatcgaccatgcccaagaagaagaggaaggTGTCCAATTTACTGACCGTACACCAAAATTTGCCTGCATTACCGGTCGATGCAACGAGTGATGAGGTTCGCAAGAACCTGATGGACATGTTCAGGGATCGCCAGGCGTTTTCTGAGCATACCTGGAAAATGCTTCTGTCCGTTTGCCGGTCGTGGGCGGCATG

 

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
35249 TGA ATC TGG CTT CCT CAA CTA TTC T Internal Positive Control Forward A
35559 TCA CGG TTT CAA TTC TCC TC Transgene Forward A
35560 GTG TAC GGT CAG TAA ATT GGA CA Transgene Reverse A
35561 Fluorophore-1 AGG GGC GGA GCC AGC AC Quencher-1 Tg Probe
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
TmoIMR0105 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 IC Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
35249 0.40 uM
35559 0.40 uM
35560 0.40 uM
oIMR3580 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.