Stock No: 012257
Protocol 19149: Standard PCR Assay - Generic SV40 Alternate1
Version 4.0

Notes

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

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

Transgene = ~300 bp

Internal positive control = 168  bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
21157 CTC CCA ACC CCA GAG GTA GT Internal Positive Control Forward A
21159 CTA TAG GGC CTG GGT CAG TG Internal Positive Control Reverse A
oIMR1201 GTG AAG GAA CCT TAC TTC TGT GGT G Transgene Forward A
oIMR1202 GTC CTT GGG GTC TTC TAC CTT TCT C Transgene Reverse A

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
21157 0.25 uM
21159 0.25 uM
oIMR1201 0.50 uM
oIMR1202 0.50 uM
Glycerol 6.50 %
Dye 1.00 X
Kapa 2G HS taq polym 0.03 U/ul
DNA

Strains Using This Protocol

Stock Number Strain Name
036602 B10.Cg-H2k2 H2-T18a Tg(SV40-HA)HA104Ajca/VradMmjax
012257 CB6-Tg(Tyr-TAg)BJjw/Mmjax
2 strains use this protocol