Stock No: 006494
Protocol 34877: Standard PCR Assay - Tg(HDexon1)62Gpb CAG Sizing
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

Flanking region 122bp

JR 6494 = ~120 repeats

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19129 Fluorophore CTC CTC GGC GGC TGA GGA AGC TGA GGA G
19808 Fluorophore CAG CTT GCC ATG GCG ACC CTG GAA AAG CTG

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTPS-kapa 0.26 mM
Primer 1 0.50 uM
Primer 2 0.50 uM
Glycerol 6.50 %
Dye 1.00 X
Kapa 2G HS taq polym 0.03 U/ul
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
006494 B6CBA-Tg(HDexon1)62Gpb/3J
027421 B6J.Cg-Tg(HDexon1)62Gpb/160ChdiJ
2 strains use this protocol