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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
Mutant = TAA
Wild type = TAT
amplicon = 149 bp
Sequence
CTGTTCCAGTTATAGATCTTTGTTTTAGGGTCATTACTTGGTTTAATGTT
TTTTAATGTAATTTGTATATGCTATTATAATAAGTAGAAAAAAATGTGTT
TTTTCCCTTAGAGTTTTGAGCAGACAATGCTGAGAAAAGGAGGATCATGG
ATTCAAGAAATAAATGTAACGGAAAAGAATTGGTATCCACAACATAAAAT
ACGCTA[t/a]GCTAAGAGAAAGTTAGCAGGGGCCAACCCAGCTGTTATAACTT
GGTAAGACTTGTGAATGCAGAATCAGTGTGTGTTCAAAAGTGCAAAGCAC
TTCACACACTTCTGAGCAGTATGGCACTTCACTGTGTAGATGGAGAAAGT
GACTCTTAGGGCGGCTTTACCCCTCCAAGCCCAGCCTGCAAGGACTGGGC
TCACACCCTTGTC
JAX Protocol
Protocol Primers
| Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
|---|---|---|---|---|---|---|
| 14102 | CAG ACA ATG CTG AGA AAA GGA G | Forward | A | |||
| 14103 | CTG CAT TCA CAA GTC TTA CCA AG | Reverse | A | |||
| 14104 | Fluorophore-1 | TAA AAT ACG CTA AGC TAA GAG AAA G | Quencher-1 | MUT Probe | ||
| 14105 | Fluorophore-2 | TAA AAT ACG CTA TGC TAA GAG AAA G | Quencher-2 | WT Probe |
Reaction A
| Component | Final Concentration |
|---|---|
| Kapa Probe Fast QPCR | 1.00 X |
| ddH2O | |
| 14102 | 0.40 uM |
| 14103 | 0.40 uM |
| Wt Probe | 0.15 uM |
| Mutant Probe | 0.15 uM |
| 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 | 40.0 | -- | 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 |
|---|---|
| 018018 | B10ScSn.Cg-Prkdcscid Dmdmdx/J |
| 017917 | B6.Cg-Dysfprmd Prkdcscid/J |
| 021146 | B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(CAG-FCGRT)276Dcr/DcrJ |
| 018441 | B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(FCGRT)32Dcr/DcrJ |
| 018541 | B6.Cg-Fcgrttm1Dcr Prkdcscid/DcrJ |
| 001913 | B6.Cg-Prkdcscid/SzJ |
| 031644 | B6.Cg-Tg(FCGRT)32Dcr Albem12Mvw Fcgrttm1Dcr Prkdcscid /J |
| 010816 | B6;C-Ghrhrlit Prkdcscid/BmJ |
| 030205 | C3.Cg-Prkdcscid Tg(Mt1-HGF)1Gvw/GvwJ |
| 001131 | C3SnSmn.Cg-Prkdcscid/J |
| 000666 | C57BL/10SnJ |
| 001803 | CBySmn.Cg-Prkdcscid/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 |
| 002313 | NOD.Cg-Emv30b Prkdcscid/Dvs |
| 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 |
| 037322 | NOD.Cg-Flt3em2Mvw Prkdcscid Il2rgtm1Wjl Tg(FLT3LG)7Sz Tg(IL15)1Sz/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 |
| 039051 | NOD.Cg-Ggta1tm1Jbl Prkdcscid Il2rgtm1Wjl/J |
| 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 |
| 039496 | NOD.Cg-Il6em1(IL6)Akp Flt3em1Akp Prkdcscid Il2rgtm1Wjl/AkpJ |
| 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 |
| 006609 | NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid/DvsJ |
| 036071 | NOD.Cg-Mertkem1Doi Prkdcscid/RweiJ |
| 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 |
| 039414 | NOD.Cg-Prkdcscid Glatm1Kul/MedinJ |
| 004606 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Tg(HLA-DQA1,HLA-DQB1)1Dv/SzJ |
| 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 |
| 039055 | NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/DvsJ |
| 032927 | NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl/SzJ |
| 026222 | NOD.Cg-Prkdcscid Hprt1em1Mvw Il2rgtm1Wjl/MvwJ |
| 004083 | NOD.Cg-Prkdcscid Iduatm1Clk/J |
| 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 |
| 017620 | NOD.Cg-Prkdcscid Tg(CAG-DsRed*MST)1Nagy/KupwJ |
| 017619 | NOD.Cg-Prkdcscid Tg(CAG-EGFP)1Osb/KupwJ |
| 001303 | NOD.Cg-Prkdcscid/J |
| 036917 | NOD.Cg-Prltm1.1(PRL)Rui Prkdcscid Il2rgtm1Wjl/J |
| 024099 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J |
| 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 |
| 022396 | NOR.Cg-Prkdcscid/JsdJ |
| 014543 | STOCK Hgftm1.1(HGF)Aveo Prkdcscid/J |
| 034329 | STOCK Lama2tm1Eeng Prkdcscid/BurkJ |
| 88 strains use this protocol | |