Protocol 13167: Separated PCR Assay - GAL Panel

ALR/LtJ

Stock No: 003070

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)-Il3^tm1Tyb/J
033315	B6.129S4(B6J)-Ppara^tm1Gonz/J
033903	B6.129S4(C)-C5ar1^tm1Cge/BaoluJ
026293	B6.Cg-Gt(ROSA)26Sor^tm1(LRRK2*R1441C)Djmo/J
021772	B6.MRL-Atp6v1b1^vtx/Kjn
017893	B6;CBA-Tg(Thy1-GCaMP3)6Gfng/J
028239	B6J.129(Cg)-Igs2^tm1.1(CAG-cas9*)Mmw/J
028861	B6J.129S6-Chat^tm2(cre)Lowl/MwarJ
025114	B6N.Cg-Tg(Camk2a-P2rx2/YC3.1)21Khakh/J
025975	BXD124/RwwJ
025985	BXD161/RwwJ
035247	C.Cg-Tg(K18-ACE2)2Prlmn/J
007600	C57BL/6J-Chr 10.1^PWD/Ph/ForeJ
006226	C57BL/6J-Chr 4^PWD/Ph/ForeJ
005304	C57BL/6NJ
014535	CAST.129S1(B6)-Kif18a^gcd2/JcsMmjax
000656	CBA/J
022527	D2.Cg-Terc^tm1Rdp/J
026558	FVB.129(B6)-Gt(ROSA)26Sor^tm1.1(CAG-cas9*,-EGFP)Fezh/J
021771	MRL/MpJ-Atp6v1b1^vtx/Kjn
21 strains use this protocol