Stock No: 002222
Protocol 22082: End Point Analysis Assay - Smcx/Smcy-PROBE
Version 2.0

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

Male = X/Y (Het)
Female = X/X (Wt)

For Standard PCR:  Omit probes.  Primers will produce the following:

WT (X Chromosome) = 123 bp

Mut (Y Chromosome) = 78 bp

Males will appear "het"

Females will appear "wt"

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
21115 AAA GGA ATT CTG GAG GCT GG Mutant Forward A
21116 TCT TTG GAG TGA ATG TCT GCT C Mutant Reverse A
21117 Fluorophore-1 AGC AGC ATT GAG GAG GAT GTG ACA G Quencher-1 MUT Probe
21118 Fluorophore-2 GAC TAG GTT CAT AGG CAC TGG Quencher-2 Wild type Forward A
21119 Fluorophore-3 CCG CCA AAA CTC CTT CTC TAC Quencher-3 Wild type Reverse A
21120 Fluorophore-4 CCC CAG ATG GTA CCC ACA GAA CTT G Quencher-4 WT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
21115 0.40 uM
21116 0.40 uM
21118 0.40 uM
21119 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 4.0 -- 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
030931 129S6.Cg-Pex1tm1.1Sjms/Mmjax
031827 B6(FVB)-Tlr7tm1.1Gbrt/J
032934 B6.129-Cxcr1tm1Gao/J
002222 B6.129S7-Twist1tm1Bhr/J
016590 B6.129S7-Ube3atm1Alb/J
030711 B6;129P2-Sumf1Gt(RST760)Byg/ClearJ
021334 B6;129S1-Ddx10m1J/GrsrJ
003263 B6;129S2-Cdkn1atm1Tyj/J
012836 B6;SJL-Tg(Thy1-TARDBP)4Singh/J
028975 B6N(Cg)-Ngly1tm2d(KOMP)Wtsi/LutzyJ
016894 C3.Cg-Kif1algdg/GrsrJ
028221 C57BL/6J-Ngly1em10Lutzy/J
031170 C57BL/6J-Smsem2Lutzy/J
027080 C57BL/6J-Tbk1em3Lutzy/1J
002063 C57BL/6JEi-Tg(Sry)1Ei Chr YAKR/J/EiJ
002708 C57BL/6JEi-Tg(Sry)2Ei Chr YAKR/J/EiJ
002837 C57BL/6JEi-Tg(Sry)3Ei Chr YAKR/J/EiJ
002183 C57BL/6JEi-Tg(Sry)4Ei Chr YPOS/EiJ
029582 C57BL/6N-Trpv4em1(cre/ERT2)Amc/J
005304 C57BL/6NJ
032808 C57BL/6NJ-2610507B11Rikem1(IMPC)J/Mmjax
032690 C57BL/6NJ-Asap3em1(IMPC)J/Mmjax
032699 C57BL/6NJ-Tspyl1em1(IMPC)J/Mmjax
032815 C57BL/6NJ-Aampem1(IMPC)J/Mmjax
034321 C57BL/6NJ-Aatfem1(IMPC)J/Mmjax
035153 C57BL/6NJ-Aco2em1(IMPC)J/Mmjax
034562 C57BL/6NJ-Actl6aem1(IMPC)J/Mmjax
034355 C57BL/6NJ-Actl6bem1(IMPC)J/Mmjax
034087 C57BL/6NJ-Aftphem1(IMPC)J/Mmjax
030349 C57BL/6NJ-Anapc1em1(IMPC)J/Mmjax
032229 C57BL/6NJ-Ankrd52em1(IMPC)J/Mmjax
032819 C57BL/6NJ-Arglu1em1(IMPC)J/Mmjax
031989 C57BL/6NJ-Arhgap21em1(IMPC)J/Mmjax
034375 C57BL/6NJ-Atp6v1fem1(IMPC)J/Mmjax
033960 C57BL/6NJ-Banpem1(IMPC)J/Mmjax
035759 C57BL/6NJ-Bicraem1(IMPC)J/Mmjax
035826 C57BL/6NJ-Bicralem1(IMPC)J/Mmjax
034353 C57BL/6NJ-Camsap1em1(IMPC)J/Mmjax
034437 C57BL/6NJ-Cct7em1(IMPC)J/Mmjax
034324 C57BL/6NJ-Cdc37em1(IMPC)J/Mmjax
033778 C57BL/6NJ-Cdc5lem1(IMPC)J/Mmjax
031318 C57BL/6NJ-Cdk17em1(IMPC)J/Mmjax
033834 C57BL/6NJ-Cfdp1em1(IMPC)J/Mmjax
032021 C57BL/6NJ-Cherpem1(IMPC)J/Mmjax
030344 C57BL/6NJ-Chmp7em1(IMPC)J/Mmjax
033693 C57BL/6NJ-Clcf1em1(IMPC)J/Mmjax
034417 C57BL/6NJ-Clec16aem1(IMPC)J/Mmjax
032957 C57BL/6NJ-Cmtr1em1(IMPC)J/Mmjax
032825 C57BL/6NJ-Cnep1r1em1(IMPC)J/Mmjax
031016 C57BL/6NJ-Cnot10em1(IMPC)J/Mmjax
034326 C57BL/6NJ-Cog8em1(IMPC)J/Mmjax
031685 C57BL/6NJ-Coq3em1(IMPC)J/Mmjax
034635 C57BL/6NJ-Cpdem1(IMPC)J/Mmjax
032761 C57BL/6NJ-Crcpem1(IMPC)J/Mmjax
033272 C57BL/6NJ-Cstf1em1(IMPC)J/Mmjax
032762 C57BL/6NJ-Cyp4f39em1(IMPC)J/Mmjax
031213 C57BL/6NJ-Dap3em1(IMPC)J/Mmjax
034083 C57BL/6NJ-Ddx31em1(IMPC)J/Mmjax
032946 C57BL/6NJ-Dhx37em1(IMPC)J/Mmjax
035025 C57BL/6NJ-Dnaaf3em1(IMPC)J/Mmjax
033866 C57BL/6NJ-Dnajb14em1(IMPC)J/Mmjax
033950 C57BL/6NJ-Dnajc2em1(IMPC)J/Mmjax
034212 C57BL/6NJ-Dzip1lem1(IMPC)J/Mmjax
034082 C57BL/6NJ-Ehbp1em1(IMPC)J/Mmjax
035167 C57BL/6NJ-Eid2bem1(IMPC)J/Mmjax
034007 C57BL/6NJ-Eif5bem1(IMPC)J/Mmjax
034322 C57BL/6NJ-Elp6em1(IMPC)J/Mmjax
031070 C57BL/6NJ-Exoc2em1(IMPC)J/Mmjax
033947 C57BL/6NJ-Exosc1em1(IMPC)J/Mmjax
034010 C57BL/6NJ-Exosc2em1(IMPC)J/Mmjax
032952 C57BL/6NJ-Faxcem1(IMPC)J/Mmjax
030192 C57BL/6NJ-Fdx2em1Murr/Murr
034221 C57BL/6NJ-Foxb2em1(IMPC)J/Mmjax
032144 C57BL/6NJ-Frmd4aem1(IMPC)J/Mmjax
036002 C57BL/6NJ-Gcn1em1(IMPC)J/Mmjax
035294 C57BL/6NJ-Gid8em1(IMPC)J/Mmjax
031116 C57BL/6NJ-Gramd1bem1(IMPC)J/Mmjax
034716 C57BL/6NJ-Gtf2h3em1(IMPC)J/Mmjax
035200 C57BL/6NJ-H2az1em1(IMPC)J/Mmjax
030731 C57BL/6NJ-Hmg20aem1(IMPC)J/Mmjax
032434 C57BL/6NJ-Hspd1em1(IMPC)J/Mmjax
032041 C57BL/6NJ-Ier3ip1em1(IMPC)J/Mmjax
033961 C57BL/6NJ-Ino80cem1(IMPC)J/Mmjax
032228 C57BL/6NJ-Ints3em1(IMPC)J/Mmjax
032235 C57BL/6NJ-Ints7em1(IMPC)J/Mmjax
034049 C57BL/6NJ-Ist1em1(IMPC)J/Mmjax
034755 C57BL/6NJ-Isy1em1(IMPC)J/Mmjax
032942 C57BL/6NJ-Kansl3em1(IMPC)J/Mmjax
033445 C57BL/6NJ-Kctd5em1(IMPC)J/Mmjax
034700 C57BL/6NJ-Kinem1(IMPC)J/Mmjax
030689 C57BL/6NJ-Kri1em1(IMPC)J/Mmjax
033815 C57BL/6NJ-Lin52em1(IMPC)J/Mmjax
034318 C57BL/6NJ-Lipmem1(IMPC)J/Mmjax
030431 C57BL/6NJ-Lpgat1em1(IMPC)J/Mmjax
033797 C57BL/6NJ-Luc7l3em1(IMPC)J/Mmjax
031886 C57BL/6NJ-Mak16em1(IMPC)J/Mmjax
031396 C57BL/6NJ-Map7d1em1(IMPC)J/Mmjax
033978 C57BL/6NJ-Mbipem1(IMPC)J/Mmjax
030128 C57BL/6NJ-Mccc2em1(IMPC)J/Mmjax
034337 C57BL/6NJ-Mecomem1(IMPC)J/Mmjax
032307 C57BL/6NJ-Mon2em1(IMPC)J/Mmjax
032943 C57BL/6NJ-Mphosph10em1(IMPC)J/Mmjax
033959 C57BL/6NJ-Mphosph8em1(IMPC)J/Mmjax
031910 C57BL/6NJ-Mpripem1(IMPC)J/Mmjax
033919 C57BL/6NJ-Mtx2em1(IMPC)J/Mmjax
032599 C57BL/6NJ-Mvdem1(IMPC)J/Mmjax
033292 C57BL/6NJ-Naa35em1(IMPC)J/Mmjax
034208 C57BL/6NJ-Ncapd3em1(IMPC)J/Mmjax
032907 C57BL/6NJ-Ndc80em1(IMPC)J/Mmjax
031127 C57BL/6NJ-Ndufa9em1(IMPC)J/Mmjax
034166 C57BL/6NJ-Noc4lem1(IMPC)J/Mmjax
031882 C57BL/6NJ-Nol9em1(IMPC)J/Mmjax
032945 C57BL/6NJ-Nufip2em1(IMPC)J/Mmjax
032140 C57BL/6NJ-Nup153em1(IMPC)J/Mmjax
033477 C57BL/6NJ-Orc2em1(IMPC)J/Mmjax
033899 C57BL/6NJ-Papolaem1(IMPC)J/Mmjax
033863 C57BL/6NJ-Pfasem1(IMPC)J/Mmjax
032589 C57BL/6NJ-Pfdn1em1(IMPC)J/Mmjax
034292 C57BL/6NJ-Pgap1em1(IMPC)J/Mmjax
032124 C57BL/6NJ-Pgm1em1(IMPC)J/Mmjax
030727 C57BL/6NJ-Phkbem1(IMPC)J/Mmjax
031380 C57BL/6NJ-Pkmyt1em1(IMPC)J/Mmjax
030819 C57BL/6NJ-Plod2em1(IMPC)J/Mmjax
031996 C57BL/6NJ-Pmpcaem1(IMPC)J/Mmjax
030726 C57BL/6NJ-Polr1cem1(IMPC)J/Mmjax
034581 C57BL/6NJ-Polr2gem1(IMPC)J/Mmjax
035199 C57BL/6NJ-Pop5em1(IMPC)J/Mmjax
034667 C57BL/6NJ-Ppa2em1(IMPC)J/Mmjax
032366 C57BL/6NJ-Ppfia1em1(IMPC)J/Mmjax
034715 C57BL/6NJ-Prebem1(IMPC)J/Mmjax
031259 C57BL/6NJ-Prim1em1(IMPC)J/Mmjax
030154 C57BL/6NJ-Prpf40aem1(IMPC)J/Mmjax
031911 C57BL/6NJ-Prrc2cem1(IMPC)J/Mmjax
034418 C57BL/6NJ-Psip1em1(IMPC)J/Mmjax
032340 C57BL/6NJ-Psmd13em1(IMPC)J/Mmjax
032231 C57BL/6NJ-Psmd2em1(IMPC)J/Mmjax
032330 C57BL/6NJ-Psmd6em1(IMPC)J/Mmjax
033799 C57BL/6NJ-Pus7em1(IMPC)J/Mmjax
031031 C57BL/6NJ-Rab10em1(IMPC)J/Mmjax
030906 C57BL/6NJ-Rab8aem1(IMPC)J/Mmjax
035070 C57BL/6NJ-Rbm45em1(IMPC)J/Mmjax
035297 C57BL/6NJ-Rexo1em1(IMPC)J/Mmjax
034714 C57BL/6NJ-Rfc5em1(IMPC)J/Mmjax
031296 C57BL/6NJ-Riok1em1(IMPC)J/Mmjax
032142 C57BL/6NJ-Rnf40em1(IMPC)J/Mmjax
032445 C57BL/6NJ-Rprd2em1(IMPC)J/Mmjax
033635 C57BL/6NJ-Rsu1em1(IMPC)J/Mmjax
030907 C57BL/6NJ-Rxylt1em1(IMPC)J/Mmjax
031168 C57BL/6NJ-Sap18em1(IMPC)J/Mmjax
034515 C57BL/6NJ-Scocem1(IMPC)J/Mmjax
030871 C57BL/6NJ-Sdhaf2em1(IMPC)J/Mmjax
032350 C57BL/6NJ-Sec62em1(IMPC)J/Mmjax
032171 C57BL/6NJ-Sf3b3em1(IMPC)J/Mmjax
034816 C57BL/6NJ-Skor1em1(IMPC)J/Mmjax
032960 C57BL/6NJ-Slain2em1(IMPC)J/Mmjax
032274 C57BL/6NJ-Slc39a10em1(IMPC)J/Mmjax
034357 C57BL/6NJ-Smarcc2em1(IMPC)J/Mmjax
034393 C57BL/6NJ-Smarce1em1(IMPC)J/Mmjax
034509 C57BL/6NJ-Snx12em1(IMPC)J/Mmjax
033009 C57BL/6NJ-Spc25em1(IMPC)J/Mmjax
031480 C57BL/6NJ-Srbd1em1(IMPC)J/Mmjax
031641 C57BL/6NJ-Srsf6em1(IMPC)J/Mmjax
034395 C57BL/6NJ-Ss18em1(IMPC)J/Mmjax
032339 C57BL/6NJ-Stt3bem1(IMPC)J/Mmjax
030915 C57BL/6NJ-Taf13em1(IMPC)J/Mmjax
034012 C57BL/6NJ-Taf5em1(IMPC)J/Mmjax
034011 C57BL/6NJ-Tbc1d32em1(IMPC)J/Mmjax
035107 C57BL/6NJ-Tbccem1(IMPC)J/Mmjax
032128 C57BL/6NJ-Tedc2em1(IMPC)J/Mmjax
033038 C57BL/6NJ-Tex36em1(IMPC)J/Mmjax
031076 C57BL/6NJ-Thap1em1(IMPC)J/Mmjax
035747 C57BL/6NJ-Thap12em1(IMPC)J/Mmjax
034865 C57BL/6NJ-Thoc3em1(IMPC)J/Mmjax
031044 C57BL/6NJ-Tm2d3em1(IMPC)J/Mmjax
032812 C57BL/6NJ-Tmem131em1(IMPC)J/Mmjax
033848 C57BL/6NJ-Tmem147em1(IMPC)J/Mmjax
034078 C57BL/6NJ-Tmem67em1(IMPC)J/Mmjax
032598 C57BL/6NJ-Tmieem1(IMPC)J/Mmjax
031169 C57BL/6NJ-Tnnc1em1(IMPC)J/Mmjax
032809 C57BL/6NJ-Togaram1em1(IMPC)J/Mmjax
031849 C57BL/6NJ-Tomm20em1(IMPC)J/Mmjax
030914 C57BL/6NJ-Tpk1em1(IMPC)J/Mmjax
034440 C57BL/6NJ-Tpx2em1(IMPC)J/Mmjax
031415 C57BL/6NJ-Trfem1(IMPC)J/Mmjax
034561 C57BL/6NJ-Tsen54em1(IMPC)J/Mmjax
034412 C57BL/6NJ-Ttc26em1(IMPC)J/Mmjax
031993 C57BL/6NJ-Ttc7em1(IMPC)J/Mmjax
032448 C57BL/6NJ-Ttc7bem1(IMPC)J/Mmjax
034438 C57BL/6NJ-Ttkem1(IMPC)J/Mmjax
034637 C57BL/6NJ-U2af2em1(IMPC)J/Mmjax
033800 C57BL/6NJ-U2surpem1(IMPC)J/Mmjax
031573 C57BL/6NJ-Ube2iem1(IMPC)J/Mmjax
030618 C57BL/6NJ-Ubiad1em1(IMPC)J/Mmjax
034804 C57BL/6NJ-Urm1em1(IMPC)J/Mmjax
032355 C57BL/6NJ-Utp20em1(IMPC)J/Mmjax
031039 C57BL/6NJ-Utp4em1(IMPC)J/Mmjax
032958 C57BL/6NJ-Vps26cem1(IMPC)J/Mmjax
034722 C57BL/6NJ-Vps36em1(IMPC)J/Mmjax
031108 C57BL/6NJ-Washc4em1(IMPC)J/Mmjax
033040 C57BL/6NJ-Wdr18em1(IMPC)J/Mmjax
034513 C57BL/6NJ-Wdr25em1(IMPC)J/Mmjax
033597 C57BL/6NJ-Wdr73em1(IMPC)J/Mmjax
033079 C57BL/6NJ-Wdr74em1(IMPC)J/Mmjax
035024 C57BL/6NJ-Wdr91em1(IMPC)J/Mmjax
032443 C57BL/6NJ-Xylt1em1(IMPC)J/Mmjax
031117 C57BL/6NJ-Yeats2em1(IMPC)J/Mmjax
034297 C57BL/6NJ-Ykt6em1(IMPC)J/Mmjax
034717 C57BL/6NJ-Zbtb34em1(IMPC)J/Mmjax
032447 C57BL/6NJ-Zc3h13em1(IMPC)J/Mmjax
035071 C57BL/6NJ-Zfhx4em1(IMPC)J/Mmjax
031035 C57BL/6NJ-Zfp407em1(IMPC)J/Mmjax
033747 C57BL/6NJ-Zfp638em1(IMPC)J/Mmjax
033578 C57BL/6NJ-Zfp652em1(IMPC)J/Mmjax
031022 C57BL/6NJ-Zfp706em1(IMPC)J/Mmjax
030432 C57BL/6NJ-Zfp827em1(IMPC)J/Mmjax
034017 C57BL/6NJ-Zmym2em1(IMPC)J/Mmjax
033981 C57BL/6NJ-Znhit1em1(IMPC)J/Mmjax
034809 C57BL/6NJ-Znhit2em1(IMPC)J/Mmjax
009617 NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid Il2rgtm1Wjl/SzJ
219 strains use this protocol