Protocol 21973: End Point Analysis Assay - H2<d> rs50413633-Alt 1-EP

BALB/cJ

Stock No: 000651

Version 2.0

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

Mut=T/T (H2<d>)

WT= A/A (other haplotypes)

Sequence

Aaatactgatcatgcaatggctgggc(A/T)gaagagagaata

gggtgGGACATCTGCCAATCA

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
21926	Fluorophore-1	ATG GCT GGG CAG AAG AGA G	Quencher-1	WT Probe
21928	Fluorophore-2	ATG GCT GGG CTG AAG AGA G	Quencher-2	MUT Probe
22202		CCA AGA ACC TCC TCT CTC ATG T		Forward	A
22203		GCA GAT GTC CCA CCC TAT TC		Reverse	A

Reaction A

Component	Final Concentration
Kapa Probe Fast QPCR	1.00 X
ddH2O
22202	0.40 uM
22203	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
027528	B10.Cg-H2^d Tg(Alb1-HBV)66Bri/ChiJ
000462	B10.D2-H2^d/n2SnJ
000461	B10.D2-Hc^0 H2^d H2-T18^c/oSnJ
000463	B10.D2-Hc^1 H2^d H2-T18^c/nSnJ
000651	BALB/cJ
002845	C.B-H2^b Tg(H2-D^d)D8Gja/LilJ
000666	C57BL/10SnJ
004353	C57BL/6-Tg(UBC-GFP)30Scha/J
007076	CByJ.B6-Tg(UBC-GFP)30Scha H2^b/J
034677	CByJ.B6-Tg(UBC-GFP)30Scha/J
028062	STOCK Ptprc^a Tcrb^Ln1Bdb Tcra^Ln1Bdb H2^d/J
11 strains use this protocol