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Stock No: 003169
Protocol 23837: Standard PCR Assay - Il2rg<tm1Wjl>
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
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-Rag1tm1Mom Cd47tm1Fpl Il2rgtm1Wjl/SzJ |
| 025730 | B6.129S-Rag2tm1Fwa Cd47tm1Fpl Il2rgtm1Wjl/J |
| 003174 | B6.129S4-Il2rgtm1Wjl/J |
| 039001 | B6.Cg-Rag2tm1Fwa Sgshmps3a Cd47tm1Fpl Il2rgtm1Wjl/JnolJ |
| 003169 | C.129S4-Il2rgtm1Wjl/J |
| 030547 | NOD.Cg-B2mtm1Unc Prkdcscid H2dlAb1-Ea Il2rgtm1Wjl/SzJ |
| 010636 | NOD.Cg-B2mtm1Unc Prkdcscid Il2rgtm1Wjl/SzJ |
| 038279 | NOD.Cg-Capn3em5Lutzy Prkdcscid Il2rgtm1Wjl/VitaJ |
| 031900 | NOD.Cg-Cmahem1Guru Prkdcscid Il2rgtm1Wjl/GuruJ |
| 028615 | NOD.Cg-Fcgrttm1Dcr Prkdcscid Il2rgtm1Wjl Tg(FCGRT)32Dcr/J |
| 035844 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(HLA-A/H2-D/B2M)1Dvs/J |
| 035843 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J |
| 035842 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl/J |
| 033367 | NOD.Cg-Flt3em2Mvw Prkdcscid Il2rgtm1Wjl Tg(FLT3LG)7Sz/SzJ |
| 026263 | NOD.Cg-Foxn1em1Dvs Prkdcscid Il2rgtm1Wjl/J |
| 029819 | NOD.Cg-Gcgem1Dvs Prkdcscid Il2rgtm1Wjl/DvsJ |
| 035959 | NOD.Cg-Gt(ROSA)26Sorem27(KRT18-ACE2)Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
| 038123 | NOD.Cg-Gt(ROSA)26Sorem28(KRT18-ACE2)Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
| 036151 | NOD.Cg-Gt(ROSA)26Sorem5Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
| 030511 | NOD.Cg-Hc1 Prkdcscid Il2rgtm1Wjl/SzJ |
| 014553 | NOD.Cg-Hgftm1.1(HGF)Aveo Prkdcscid Il2rgtm1Wjl/J |
| 035868 | NOD.Cg-Iduaem1Osp Prkdcscid Il2rgtm1Wjl/J |
| 035007 | NOD.Cg-Iduatm1Clk Prkdcscid Il2rgtm1Wjl/J |
| 026622 | NOD.Cg-KitW-41J Tyr + Prkdcscid Il2rgtm1Wjl/ThomJ |
| 026497 | NOD.Cg-KitW-41J Prkdcscid Il2rgtm1Wjl/WaskJ |
| 009617 | NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid Il2rgtm1Wjl/SzJ |
| 032474 | NOD.Cg-Prkdcscid Abcd1em2Lutzy Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ |
| 027905 | NOD.Cg-Prkdcscid Cd274tm1Shr Il2rgtm1Wjl/J |
| 030610 | NOD.Cg-Prkdcscid Cybbem1Hmal Il2rgtm1Wjl/HmalJ |
| 031566 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ |
| 026936 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv Tg(INS*)172Dvs/DvsJ |
| 026561 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/SzJ |
| 021885 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl/SzJ |
| 025216 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-Ab1g7-em1Mvw H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
| 023848 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
| 032927 | NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl/SzJ |
| 026222 | NOD.Cg-Prkdcscid Hprt1em1Mvw Il2rgtm1Wjl/MvwJ |
| 028657 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(CSF1)3Sz/J |
| 014570 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-A/H2-D/B2M)1Dvs/SzJ |
| 030890 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(IL15)1Sz/SzJ |
| 035970 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(YAC128)53Hay/JnolJ |
| 035424 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Ace2em1Lutzy/J |
| 035002 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Ace2tm1(ACE2)Dwnt/J |
| 017637 | NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ |
| 012480 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Hprt1b-m3/EshJ |
| 034859 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-cre/Esr1*)5Amc/J |
| 021937 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-EGFP)1Osb/SzJ |
| 013062 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ |
| 028654 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CSF1)3Sz/SzJ |
| 029295 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ |
| 028655 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(IL6)1Sz/SzJ |
| 027976 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(Ins2-HBEGF)6832Ugfm/SzJ |
| 017830 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(PGK1-KITLG*220)441Daw/SzJ |
| 028842 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(SERPINA1*E342K)#Slcw/SzJ |
| 005557 | NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ |
| 034569 | NOD.Cg-Prkdcscid Otcem1Bshep Il2rgtm1Wjl/J |
| 011066 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CSF2)2Ygy Tg(IL3)1Ygy Tg(KITLG)3Ygy/YgyJGckRolyJ |
| 036917 | NOD.Cg-Prltm1.1(PRL)Rui Prkdcscid Il2rgtm1Wjl/J |
| 030442 | NOD.Cg-Rag1tm1Mom Dmdmdx-4Cv Il2rgtm1Wjl/McalJ |
| 018454 | NOD.Cg-Rag1tm1Mom Fahem1Mvw Il2rgtm1Wjl/MvwJ |
| 026062 | NOD.Cg-Rag1tm1Mom Fancaem1Dvs Il2rgtm1Wjl/SzJ |
| 033127 | NOD.Cg-Rag1tm1Mom Flt3tm1Irl Mcph1Tg(HLA-A2.1)1Enge Il2rgtm1Wjl/J |
| 035855 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-A/H2-D/B2M)Dvs Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/J |
| 030533 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(SLC10A1)15Mvw/MvwJ |
| 026014 | NOD.Cg-Rag1tm1Mom KitW-41J Il2rgtm1Wjl/EavJ |
| 030443 | NOD.Cg-Rag1tm1Mom Sgcatm1Kcam Il2rgtm1Wjl/McalJ |
| 029663 | NOD.Cg-Rag1tm1Mom Dysfprmd Il2rgtm1Wjl/McalJ |
| 024099 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J |
| 017914 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/ScasJ |
| 007799 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ |
| 014568 | NOD.Cg-Rag1tm1Mom Ins2Akita Il2rgtm1Wjl/SzJ |
| 033704 | NOD.Cg-Tlr4lps-del Prkdcscid Il2rgtm1Wjl/SzJ |
| 028616 | NOD.Cg-Tg(CAG-FCGRT)276Dcr Fcgrttm1Dcr Prkdcscid Il2rgtm1Wjl/J |
| 026997 | NOD.Cg-Tg(CD68-EGFP)1Drg Prkdcscid Il2rgtm1Wjl/J |
| 030331 | NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl/SzJ |
| 012479 | NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdcscid Il2rgtm1Wjl/GckRolyJ |
| 034901 | NOD.Cg-Tg(K18-ACE2)2Prlmn Prkdcscid Il2rgtm1Wjl/J |
| 77 strains use this protocol | |