Protocol 23837: Standard PCR Assay - Il2rg<tm1Wjl>

NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ

Stock No: 005557

Version 4.2

Notes

The Il2rg gene is X-LINKED. Males with the targeted allele are hemizygous mutants

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

Mutant = ~349 bp
Heterozygote = ~269 bp and ~349 bp
Wild type = ~269 bp

Separated by gel electrophoresis on a 1.5% agarose gel.

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
oIMR5330		GTG GGT AGC CAG CTC TTC AG		Common	A
oIMR5331		CCT GGA GCT GGA CAA CAA AT		Wild type Reverse	A
oIMR7415		GCC AGA GGC CAC TTG TGT AG		Mutant Reverse	A	PGK

Reaction A

Component	Final Concentration
ddH2O
Kapa 2G HS buffer	1.30 X
MgCl2	2.60 mM
dNTP KAPA	0.26 mM
oIMR5330	0.50 uM
oIMR5331	0.50 uM
oIMR7415	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	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 (Touchdown)
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.

JAX uses a 'touchdown' cycling protocol and therefore has not calculated the optimal annealing temperature for each set of primers.

Strains Using This Protocol

Stock Number	Strain Name
032088	B6.129S-Rag1^tm1Mom Cd47^tm1Fpl Il2rg^tm1Wjl/SzJ
025730	B6.129S-Rag2^tm1Fwa Cd47^tm1Fpl Il2rg^tm1Wjl/J
003174	B6.129S4-Il2rg^tm1Wjl/J
039001	B6.Cg-Rag2^tm1Fwa Sgsh^mps3a Cd47^tm1Fpl Il2rg^tm1Wjl/JnolJ
003169	C.129S4-Il2rg^tm1Wjl/J
030547	NOD.Cg-B2m^tm1Unc Prkdc^scid H2^dlAb1-Ea Il2rg^tm1Wjl/SzJ
010636	NOD.Cg-B2m^tm1Unc Prkdc^scid Il2rg^tm1Wjl/SzJ
038279	NOD.Cg-Capn3^em5Lutzy Prkdc^scid Il2rg^tm1Wjl/VitaJ
031900	NOD.Cg-Cmah^em1Guru Prkdc^scid Il2rg^tm1Wjl/GuruJ
028615	NOD.Cg-Fcgrt^tm1Dcr Prkdc^scid Il2rg^tm1Wjl Tg(FCGRT)32Dcr/J
035844	NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(HLA-A/H2-D/B2M)1Dvs/J
035843	NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J
035842	NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/J
039497	NOD.Cg-Flt3^em1Akp Prkdc^scid Tslp^em3(TSLP)Akp Il2rg^tm1Wjl/AkpJ
037322	NOD.Cg-Flt3^em2Mvw Prkdc^scid Il2rg^tm1Wjl Tg(FLT3LG)7Sz Tg(IL15)1Sz/J
033367	NOD.Cg-Flt3^em2Mvw Prkdc^scid Il2rg^tm1Wjl Tg(FLT3LG)7Sz/SzJ
026263	NOD.Cg-Foxn1^em1Dvs Prkdc^scid Il2rg^tm1Wjl/J
029819	NOD.Cg-Gcg^em1Dvs Prkdc^scid Il2rg^tm1Wjl/DvsJ
039051	NOD.Cg-Ggta1^tm1Jbl Prkdc^scid Il2rg^tm1Wjl/J
035959	NOD.Cg-Gt(ROSA)26Sor^em27(KRT18-ACE2)Mvw Prkdc^scid Il2rg^tm1Wjl/MvwJ
038123	NOD.Cg-Gt(ROSA)26Sor^em28(KRT18-ACE2)Mvw Prkdc^scid Il2rg^tm1Wjl/MvwJ
036151	NOD.Cg-Gt(ROSA)26Sor^em5Mvw Prkdc^scid Il2rg^tm1Wjl/MvwJ
030511	NOD.Cg-Hc^1 Prkdc^scid Il2rg^tm1Wjl/SzJ
014553	NOD.Cg-Hgf^tm1.1(HGF)Aveo Prkdc^scid Il2rg^tm1Wjl/J
035868	NOD.Cg-Idua^em1Osp Prkdc^scid Il2rg^tm1Wjl/J
035007	NOD.Cg-Idua^tm1Clk Prkdc^scid Il2rg^tm1Wjl/J
039496	NOD.Cg-Il6^em1(IL6)Akp Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/AkpJ
039498	NOD.Cg-Il6^em1(IL6)Akp Flt3^em1Akp Prkdc^scid Tslp^em3(TSLP)Akp Il2rg^tm1Wjl/AkpJ
026622	NOD.Cg-Kit^W-41J Tyr ^+ Prkdc^scid Il2rg^tm1Wjl/ThomJ
026497	NOD.Cg-Kit^W-41J Prkdc^scid Il2rg^tm1Wjl/WaskJ
009617	NOD.Cg-Mcph1^Tg(HLA-A2.1)1Enge Prkdc^scid Il2rg^tm1Wjl/SzJ
032474	NOD.Cg-Prkdc^scid Abcd1^em2Lutzy Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ
027905	NOD.Cg-Prkdc^scid Cd274^tm1Shr Il2rg^tm1Wjl/J
030610	NOD.Cg-Prkdc^scid Cybb^em1Hmal Il2rg^tm1Wjl/HmalJ
031566	NOD.Cg-Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ
026936	NOD.Cg-Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv Tg(INS*)172Dvs/DvsJ
026561	NOD.Cg-Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/SzJ
021885	NOD.Cg-Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl/SzJ
025216	NOD.Cg-Prkdc^scid H2-K1^b-tm1Bpe H2-Ab1^g7-em1Mvw H2-D1^b-tm1Bpe Il2rg^tm1Wjl/SzJ
023848	NOD.Cg-Prkdc^scid H2-K1^b-tm1Bpe H2-D1^b-tm1Bpe Il2rg^tm1Wjl/SzJ
039055	NOD.Cg-Prkdc^scid H2-K1^d-em1Dvs H2-Ab1^g7-em1Dvs H2-D1^b-em5Dvs Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/DvsJ
032927	NOD.Cg-Prkdc^scid H2-K1^d-em1Dvs H2-Ab1^g7-em1Dvs H2-D1^b-em5Dvs Il2rg^tm1Wjl/SzJ
026222	NOD.Cg-Prkdc^scid Hprt1^em1Mvw Il2rg^tm1Wjl/MvwJ
028657	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(CSF1)3Sz/J
014570	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(HLA-A/H2-D/B2M)1Dvs/SzJ
030890	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(IL15)1Sz/SzJ
035970	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(YAC128)53Hay/JnolJ
035424	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Ace2^em1Lutzy/J
035002	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Ace2^tm1(ACE2)Dwnt/J
017637	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl H2-Ab1^b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ
012480	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Hprt1^b-m3/EshJ
034859	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CAG-cre/Esr1*)5Amc/J
021937	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CAG-EGFP)1Osb/SzJ
013062	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ
028654	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CSF1)3Sz/SzJ
029295	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ
028655	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(IL6)1Sz/SzJ
027976	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(Ins2-HBEGF)6832Ugfm/SzJ
017830	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(PGK1-KITLG*220)441Daw/SzJ
028842	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(SERPINA1*E342K)#Slcw/SzJ
005557	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
034569	NOD.Cg-Prkdc^scid Otc^em1Bshep Il2rg^tm1Wjl/J
011066	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CSF2)2Ygy Tg(IL3)1Ygy Tg(KITLG)3Ygy/YgyJGckRolyJ
036917	NOD.Cg-Prl^tm1.1(PRL)Rui Prkdc^scid Il2rg^tm1Wjl/J
030442	NOD.Cg-Rag1^tm1Mom Dmd^mdx-4Cv Il2rg^tm1Wjl/McalJ
018454	NOD.Cg-Rag1^tm1Mom Fah^em1Mvw Il2rg^tm1Wjl/MvwJ
026062	NOD.Cg-Rag1^tm1Mom Fanca^em1Dvs Il2rg^tm1Wjl/SzJ
033127	NOD.Cg-Rag1^tm1Mom Flt3^tm1Irl Mcph1^Tg(HLA-A2.1)1Enge Il2rg^tm1Wjl/J
035855	NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(HLA-A/H2-D/B2M)Dvs Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/J
030533	NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(SLC10A1)15Mvw/MvwJ
026014	NOD.Cg-Rag1^tm1Mom Kit^W-41J Il2rg^tm1Wjl/EavJ
030443	NOD.Cg-Rag1^tm1Mom Sgca^tm1Kcam Il2rg^tm1Wjl/McalJ
029663	NOD.Cg-Rag1^tm1Mom Dysf^prmd Il2rg^tm1Wjl/McalJ
024099	NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J
017914	NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/ScasJ
007799	NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ
014568	NOD.Cg-Rag1^tm1Mom Ins2^Akita Il2rg^tm1Wjl/SzJ
037275	NOD.Cg-Rag2^em8Lutzy H2-K1^b-tm1Bpe H2-Ab1^g7-em1Mvw H2-D1^b-tm1Bpe Il2rg^tm1Wjl/J
037276	NOD.Cg-Rag2^em8Lutzy Il2rg^tm1Wjl/J
033704	NOD.Cg-Tlr4^lps-del Prkdc^scid Il2rg^tm1Wjl/SzJ
028616	NOD.Cg-Tg(CAG-FCGRT)276Dcr Fcgrt^tm1Dcr Prkdc^scid Il2rg^tm1Wjl/J
026997	NOD.Cg-Tg(CD68-EGFP)1Drg Prkdc^scid Il2rg^tm1Wjl/J
030331	NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl/SzJ
012479	NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdc^scid Il2rg^tm1Wjl/GckRolyJ
034901	NOD.Cg-Tg(K18-ACE2)2Prlmn Prkdc^scid Il2rg^tm1Wjl/J
85 strains use this protocol