Protocol 17937: QPCR Assay - Tg(HLA-A)-1

NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ

Stock No: 029295

Version 4.0

Notes

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
oIMR1332		TTC TCC CTC TCC CAA CCT ATG TAG		Transgene
oIMR1544		CAC GTG GGC TCC AGC ATT		Internal Positive Control Forward
oIMR3580		TCA CCA GTC ATT TCT GCC TTT G		Internal Positive Control Reverse
oIMR4311		CGA CGA CAC TGA TTG GCT TCT		Transgene
TmoIMR0017	Fluorophore-1	TCA CTC CCA TTG GGT GTC GGG TTT	Quencher-1	Tg Probe
TmoIMR0105	Fluorophore-2	CCA ATG GTC GGG CAC TGC TCA A	Quencher-2	IC Probe

Strains Using This Protocol

Stock Number	Strain Name
029295	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ
033127	NOD.Cg-Rag1^tm1Mom Flt3^tm1Irl Mcph1^Tg(HLA-A2.1)1Enge Il2rg^tm1Wjl/J
2 strains use this protocol