Stock No: 000461
Protocol 21973: End Point Analysis Assay - H2<d> rs50413633-Alt 1-EP
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 4.0 -- 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-H2d Tg(Alb1-HBV)66Bri/ChiJ
000462 B10.D2-H2d/n2SnJ
000461 B10.D2-Hc0 H2d H2-T18c/oSnJ
000463 B10.D2-Hc1 H2d H2-T18c/nSnJ
000651 BALB/cJ
002845 C.B-H2b Tg(H2-Dd)D8Gja/LilJ
000666 C57BL/10SnJ
004353 C57BL/6-Tg(UBC-GFP)30Scha/J
007076 CByJ.B6-Tg(UBC-GFP)30Scha H2b/J
028062 STOCK Ptprca TcrbLn1Bdb TcraLn1Bdb H2d/J
10 strains use this protocol