Protocol 34111: Standard PCR Assay - c9ORF72-3'
Version 1.0

Notes

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

Genotyping by PCR is performed to determine the absence or presence of the transgene. This strain has some variability in repeat size, therefore Southern blotting is performed to accurately determine the size, in kilobases, of the repeat within the transgene.

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 = 348 bp

Internal positive control = 168 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
15967 TGT TTG CCT GCA ATA GGC Transgene Forward
15968 TGA TGG GAA AGC TAT TAT GAC C Transgene Reverse
21157 CTC CCA ACC CCA GAG GTA GT Internal Positive Control Forward
21159 CTA TAG GGC CTG GGT CAG TG Internal Positive Control Reverse

Strains Using This Protocol

Stock Number Strain Name
023099 C57BL/6J-Tg(C9orf72_i3)112Lutzy/J
036633 FVB/NJ-Tg(C9orf72_i3)101Lutzy/J
2 strains use this protocol