Protocol 22146: QPCR Assay - Tg(CYP1A1,CYP1A2)1Dwn
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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR1544 CAC GTG GGC TCC AGC ATT Internal Positive Control Forward A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
oIMR8522 AGA CAC TGA TCT GGC TGC AGT Transgene Forward A
oIMR8523 CAG AGG CAA GTC CAG GGT AG Transgene Reverse A
TmoIMR0105 Fluorophore-1 CCA ATG GTC GGG CAC TGC TCA A Quencher-1 IC Probe
TmoIMR0107 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 Tg Probe
TmoIMR0124 Fluorophore-3 TAT CTG GGC TGT GGG CAA GCC TAA G Quencher-3 Tg Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
oIMR1544 0.40 uM
oIMR3580 0.40 uM
oIMR8522 0.40 uM
oIMR8523 0.40 uM
Tg Probe 0.15 uM
IC Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 -- repeat steps 2-3 for 40 cycles
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.