Stock No: 005584
Protocol 36217: Probe Assay - Tg(Prrx1-cre)1Cjt Probe
Version 1.0

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=  145 bp

IPC = 132 bp

Sequence

Tg Sequence:

 tgttggcaaaggggttttcttaatcagactgttttttcggtccgaggaaaaggaggtaggagattgtgatggagaaagggggtctgtaaaacgtcaggcgCCGCACAAAGGCTTTGCCACGTAATTACAGGCTCCTATTAAGTGGAGATCCCCCGGGCTGCAGGAATTCGCCGCCACCATGCCCAAGAAGAAGAGGAAGGtgtccaatttactgaccgtacaccaaaatttgc

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
33394 AGG GGG TCT GTA AAA CGT CA Transgene Forward A Prrx1
35249 TGA ATC TGG CTT CCT CAA CTA TTC T Internal Positive Control Forward A
48147 GTA CGG TCA GTA AAT TGG ACA CC Transgene Reverse A Cre
48148 Fluorophore-1 CGC ACA AAG GCT TTG CCA C 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
33394 0.40 uM
35249 0.40 uM
48147 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

Stock Number Strain Name
005584 B6.Cg-Tg(Prrx1-cre)1Cjt/J
033173 B6N.Cg-Tg(Prrx1-cre)1Cjt/ErenJ
2 strains use this protocol