Stock No: 003303
Protocol 30013: QPCR Assay - Tg(DO11.10)10Dlo-Tcra qPCR
Version 7.0


Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler
480. Use an appropriate instrument specific Fluorophore/Quencher combination.
The transgene genotype is determined by comparing ΔCt values of each unknown
sample against known homozygous and hemizygous controls, using appropriate
endogenous references.
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
oIMR1811 AAC GCT TCT CCC TGC ACA TC Transgene Forward A
oIMR1812 AGG ACT TGC AGC ACA GAA GTA CAT Transgene Reverse A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
TmoIMR0028 Fluorophore-1 CAG ACA CCC AGC CTG GAG ACT CAG C Quencher-1 Tg Probe
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
oIMR1544 0.40 uM
oIMR1811 0.40 uM
oIMR1812 0.40 uM
oIMR3580 0.40 uM
Tg Probe 0.15 uM
IC Probe 0.15 uM


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

Stock Number Strain Name
003147 B10.D2-Hc1 H2d H2-T18c/nSnJ-Tg(DO11.10)10Dlo/J
030666 C.Cg-Rag1tm1Mom Tg(DO11.10)10Dlo/J
036332 C.Cg-Thy1a Tg(DO11.10)10Dlo/AkhoAbhmJ
003303 C.Cg-Tg(DO11.10)10Dlo/J
4 strains use this protocol