Stock No: 003225
Protocol 13167: Separated PCR Assay - GAL Panel
Version 1.0

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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.00
MgCl2 2.00
dNTPS-kapa 0.20
Primer 1 0.50
Primer 2 0.50
10x Loading Dye 0.50
Kapa 2G HS taq polym 0.01
DNA

Cycling

Step Temp °C Time Note
1 94.0 --
2 94.0 --
3 65.0 -- -0.5 C per cycle decrease
4 68.0 --
5 -- repeat steps 2-4 for 10 cycles
6 94.0 --
7 60.0 --
8 72.0 --
9 -- repeat steps 6-8 for 28 cycles
10 72.0 --
11 10.0 -- hold
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
003070 ALR/LtJ
026277 B6.129S2(B10)-Il3tm1Tyb/J
033903 B6.129S4(C)-C5ar1tm1Cge/BaoluJ
033315 B6.129S4-Pparatm1Gonz/J
026293 B6.Cg-Gt(ROSA)26Sortm1(LRRK2*R1441C)Djmo/J
028276 B6.Cg-Pdcd1tm1.1Shr/J
021772 B6.MRL-Atp6v1b1vtx/Kjn
017893 B6;CBA-Tg(Thy1-GCaMP3)6Gfng/J
028239 B6J.129(Cg)-Igs2tm1.1(CAG-cas9*)Mmw/J
028863 B6J.129S6(FVB)-Slc17a6tm2(cre)Lowl/MwarJ
028861 B6J.129S6-Chattm2(cre)Lowl/MwarJ
025114 B6N.Cg-Tg(Camk2a-P2rx2/YC3.1)21Khakh/J
000651 BALB/cJ
025975 BXD124/RwwJ
025985 BXD161/RwwJ
003225 BXD36/TyJ
007600 C57BL/6J-Chr 10.1PWD/Ph/ForeJ
006226 C57BL/6J-Chr 4PWD/Ph/ForeJ
005304 C57BL/6NJ
014535 CAST.129S1(B6)-Kif18agcd2/JcsMmjax
000656 CBA/J
022527 D2.Cg-Terctm1Rdp/J
026558 FVB.129(B6)-Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J
021771 MRL/MpJ-Atp6v1b1vtx/Kjn
24 strains use this protocol