Protocol 30795: Probe Assay - Generic Neo
Version 3.0

Notes

PLEASE NOTE: This assay will NOT distinguish heterozygous from homozygous for anything including Tcrd<tm1Mom> mutant mice. DSW 9/13/17

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


Tg = 78 bp

IPC = 74 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
32403 Fluorophore-1 CGG CGA ATG GGC TGA C Quencher-1 Tg Probe
35195 GAG CGG CGA TAC CGT AAA G Transgene Reverse A
oIMR1544 CAC GTG GGC TCC AGC ATT Internal Positive Control Forward A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
oIMR6773 TGG CTA CCC GTG ATA TTG CT Transgene Forward A
TmoIMR0105 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 IC Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
35195 0.40 uM
oIMR1544 0.40 uM
oIMR3580 0.40 uM
oIMR6773 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
025451 129-Iqgap1tm1Aber/VsJ
004166 129-Itgb5tm1Des/J
017631 129-Nrarptm1Grid/J
012560 129-Zic4tm2Kjmi/J
024906 129-Dp(16Tiam1-Il10rb)8Yey/J
037010 129;B6-Fmodtm1Aol/SooJ
028028 129S(B6N)-Cdk1tm1Eddy/J
024634 129S(Cg)-Tgfbr2tm1.1Hcd/J
007664 129S-Efnb1tm1Sor/J
024525 129S-Gnai3tm1Lbi Gnai1tm1Lbi/Mmjax
008668 129S-Hba-a1tm1Led/J
007005 129S-Scg5tm1Led/J
018424 129S-Slc26a4tm1Egr/AjgJ
030769 129S/Sv-Ccn2tm1Mae/J
035806 129S4/SvJae-Fgfr2tm6.1Sor/Mmjax
016264 129S6-Fmn2tm1Led/RwwJ
005032 B6 x B6Ei.129(Cg)-Bmp8btm1Blh/EiJ
037318 B6(Cg)-H3c6em1Sjes/J
029290 B6(Cg)-Sgtatm1.1Drmcr/J
021072 B6(Cg)-Slc6a8tm1.2Clar/J
021828 B6(SJL)-Lrrk2tm3.1Mjff/J
025930 B6(SJL)-Pik3cbtm1.1Jdo/J
028635 B6.129(Cg)-Cntnap2tm2Pele/J
023313 B6.129(Cg)-Homer2tm1Mhd/PfwJ
008320 B6.129(Cg)-Leprtm2(cre)Rck/J
037951 B6.129(Cg)-Wapltm1.1Kpfe/J
029799 B6.129-Ank3tm1Bnt/J
023853 B6.129-Chgatm1Gnh/J
004276 B6.129-Figntm1Frk/Frk
003558 B6.129-Grm5tm1Rod/J
037698 B6.129-Gt(ROSA)26Sortm13.1(CAG-HRP)Luo/J
030952 B6.129-Gt(ROSA)26Sortm5.1(CAG-Sun1/sfGFP)Nat/MmbeJ
038594 B6.129-H3f3atm1Hjf/J
005819 B6.129-Insl5tm1Dgen/J
034643 B6.129-Irgm1tm1Gat/J
028890 B6.129-Ncor1tm1Rsd/J
028891 B6.129-Ncor2tm1Rsd/J
037191 B6.129-Nefhtm2Dwc Nefmtm1Mvr/J
022996 B6.129-Nox4tm1Kkr/J
025633 B6.129-Pdk4tm1Rhar/J
028765 B6.129-Pfdn1Gt(GTR1.3)1Rul/WkhnJ
003870 B6.129-Plin1tm1Chan/J
028486 B6.129-Ppoxtm1Had/HadLlp
030664 B6.129-Prdx6tm1Abf/Mmjax
002725 B6.129-Ptprctm1Holm/J
029902 B6.129-Ptprctm1Mak/Mmjax
034226 B6.129-Rab25tm1Jrgo/J
032321 B6.129-Rsad2tm1Kchc/CresJ
006833 B6.129-Sgcbtm1Kcam/2J
023309 B6.129-Stat2tm1Shnd/J
019375 B6.129-Ttc9tm1Vcl/Mmjax
010725 B6.129-Ugt1tm1Rhtu/J
036108 B6.129P2(C)-Cnr1tm1.1Ltz/J
030985 B6.129P2(Cg)-Aph1atm1Bdes Aph1btm1Bdes Aph1ctm1Bdes/J
029253 B6.129P2(Cg)-Card6tm1.1Aldu/MakJ
022364 B6.129P2(SJL)-Ntrk3tm1Ddg/J
022499 B6.129P2-Adora2btm1Till/J
032260 B6.129P2-Apaf1tm1Mak/Mmjax
023726 B6.129P2-C1qatm1.1Tenn/J
029905 B6.129P2-Ctla4tm1Mak/Mmjax
005814 B6.129P2-Grm1tm1Dgen/J
002204 B6.129P2-Hbbtm1Unc/J
031834 B6.129P2-Irf4tm1Mak/J
024630 B6.129P2-Lcn2tm1Aade/AkiJ
029904 B6.129P2-Msh2tm1Mak/Mmjax
034729 B6.129P2-Myliptm1Pton/J
032293 B6.129P2-Rhbdf2tm1Mak/Mmjax
005845 B6.129P2-Thbs4tm1Dgen/J
034595 B6.129S-Capn1tm1Ahc/J
002729 B6.129S-Cd74tm1Liz/J
031661 B6.129S-Chattm1(cre)Lowl/MwarJ
036668 B6.129S-Grprtm2(icre)Zfc/J
002274 B6.129S-Itga5tm1Hyn/J
027187 B6.129S-Lnpeptm1Ske/J
023209 B6.129S-Sfrp1tm1Jrbn/PcmaJ
007015 B6.129S1-Gadd45gtm1Flv/J
037386 B6.129S1-Gnat3tm1Rfm/J
035475 B6.129S1-Ifi30tm1Cres/J
038977 B6.129S1-Nfkb2tm2Brv/AlexJ
030536 B6.129S1-Tfe3tm1Est/Mmjax
021055 B6.129S2(Cg)-Bdnftm1Krj/J
005027 B6.129S2-Il10rbtm1Agt/J
030087 B6.129S4(C)-Miftm1Dvd/MgadJ
006955 B6.129S4(FVB)-Insrtm1Khn/J
004163 B6.129S4-Cdk5r1tm1Lht/J
002781 B6.129S4-Cdkn1btm1Mlf/J
004185 B6.129S4-Soat2tm1Far/J
037536 B6.129S4-Sypl2tm1Hta/J
010672 B6.129S4-Tnfrsf11btm1Eac/J
018648 B6.129S6(Cg)-Itgb2tm1.1Mskim/J
031499 B6.129S6-Acer3tm1Cmao/Mmjax
003742 B6.129S6-Cdh6tm1Sma/J
017975 B6.129S6-Dag1tm5Kcam/J
030639 B6.129S6-Ece2tm1Ywa/J
018164 B6.129S6-Pdcd4tm1Yhcn/J
005623 B6.129S6-Shhtm2(cre/ERT2)Cjt/J
025929 B6.129S7(Cg)-Pik3catm1Jdo/J
008683 B6.129S7-Cacna1atm3Hzo/J
003312 B6.129S7-Ngftm1Gne/J
022767 B6.129S7-Slc7a2tm1Clm/LellJ
002277 B6.129S7-Srctm1Sor/J
002222 B6.129S7-Twist1tm1Bhr/J
028953 B6.129X1-Id4tm1Mais/Mmjax
038214 B6.C(Cg)-Gt(ROSA)26Sortm1(Stat6*)Voeh/J
035514 B6.Cg-Cd19tm1(cre)Cgn Ikzf3tm1.1Cjwu/J
036705 B6.Cg-Cd80tm1Shr Cd86tm2Shr Tg(Cd80)1Hode/J
038176 B6.Cg-Gt(ROSA)26Sortm1(CAG-APOBEC3B)Rshar/J
038190 B6.Cg-Gt(ROSA)26Sortm1(CAG-Bcl2)Zhu/J
030743 B6.Cg-Gt(ROSA)26Sortm1(CAG-Pycard/mCitrine*,-CD2*)Dtg/J
031279 B6.Cg-Gt(ROSA)26Sortm1(tetO-Yap1*,-GFP*)Stang/J
006513 B6.Cg-Ighmbp2nmd-2J Tg(Ttn-Ighmbp2)45Cx/Cx
031842 B6.Cg-Irak1tm1Jth/CpasJ
024469 B6.Cg-Lrrk2tm3.1Mjfa/J
034191 B6.Cg-Mir21atm1Yoli/RthsnJ
027748 B6.Cg-Mybl1tm1Epr/J
030544 B6.Cg-Npy1rtm1.1(cre/GFP)Rpa/J
029031 B6.Cg-Pabpc1ltm1.1Esel/J
036415 B6.Cg-Sh2b1em3Ccsu/J
013717 B6.Cg-Slc20a1tm1.2Cmg/J
012627 B6.Cg-Stard4tm1.1Bres/J
034601 B6.Cg-Tgfb1tm1Mki/J
033705 B6.Cg-Tg(Lck-RHO/ADRA1,-lacZ)B22-1Mik/J
021614 B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J
008135 B6.Cg-Tg(THY1-SNCA*A53T)M53Sud/J
035020 B6.D1-Ksr2tm1Rlw/J
036551 B6.FVB(Cg)-Tg(Pmch-cre)1Lowl/J
033109 B6;129-Cdh2tm1(cre/ERT)Linli/J
038489 B6;129-Ces1dtm2Reli/J
011082 B6;129-Edn2tm1.1Nat/J
003295 B6;129-Ednrbtm1Ywa/J
027583 B6;129-Esyt1tm1Sud Esyt2tm1Sud/J
012820 B6;129-Fzd1tm1.1Nat/J
010948 B6;129-Gabra1tm4Geh/J
024649 B6;129-Gdf7tm1Kng/J
036087 B6;129-Gt(ROSA)26Sortm3Cos/J
021039 B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J
012894 B6;129-Macf1tm1Efu/J
006904 B6;129-Msctm1Eno/J
032689 B6;129-Nefhtm2Dwc/J
006378 B6;129-Nrxn2tm3Sud/J
011081 B6;129-Osmrtm1.1Nat/J
012653 B6;129-Pax7tm1.1Fan/J
003255 B6;129-Plp1tm1Kan/J
034559 B6;129-Prdm2tm1Shg/J
025740 B6;129-Prg4tm2Mawa/J
010673 B6;129-Runx1tm3.1Spe/J
036525 B6;129-Scgb1a1tm1(icre)Fjd/J
009600 B6;129-Six2tm3(EGFP/cre/ERT2)Amc/J
018773 B6;129-Sult2b1tm1Rus/J
031104 B6;129-Tbx6tm1Pa/Mmjax
039020 B6;129-Vgftm5Srjs/AbarJ
006374 B6;129P2-Rph3atm1Sud/J
006386 B6;129P2-Syt1tm5Sud/J
026012 B6;129P2-Uts2rtm1Djbe/J
024003 B6;129S-Matn1tm1Goe/J
027581 B6;129S-Nfatc4tm1Grc/J
021968 B6;129S-Vcptm1Itl/J
035714 B6;129S1-Gt(ROSA)26Sortm1(Bcor*A)Vjba/J
003284 B6;129S1-Il1raptm1Roml/J
029304 B6;129S2-Syngap1tm2Geno/RumbJ
012623 B6;129S4-Carftm1.1Aew/J
036304 B6;129S4-Gt(ROSA)26Sortm6(CAG-tTA)Zjh/J
016835 B6;129S4-L3mbtl1tm1.1Haho/J
023343 B6;129S4-Pcmt1tm1Scl/J
003749 B6;129S5-Pak2Gt(OST104058)Lex/J
023399 B6;129S6-Ctbp2tm1Sud/J
002594 B6;129S6-Gba1tm1Nsb/J
030717 B6;129S7-Atxn1ltm2Hzo/J
033148 B6;129S7-Grprtm1Zfc/J
034237 B6;129X-Stx2tm1Dcru/J
032596 B6;129X1-Hip1rtm1Tsr/J
027102 B6;129X1-Lhcgrtm1Zmlei/J
031387 B6;CBA-Axin2em1Fstl/J
006680 B6;CBA-Tg(Olfr16*,taulacZ)19Mom/MomJ
026526 B6;SJL-Akap6tm1.1Mskf/Mmjax
007266 B6Ei.129S1(Cg)-Zfpm2tm2Sho/EiJ
037014 B6J.129P2-Pofut2Gt(RST434)Byg/BchMmjax
027410 B6J.129S1-Htttm1Mfc/190ChdiJ
370476 B6J.129S1-Htttm1Mfc/190JChdi
034731 B6N(129S4)-Srd5a3tm1c(EUCOMM)Wtsi/J
032545 B6N.129(Cg)-Cenpetm2.1Dwc/J
027971 B6N.129(Cg)-Pax6tm1Pgr/J
025330 B6N.129P2(Cg)-Igs13tm1Dolm Igs14tm1Dolm/J
021078 B6N.129S1-Mrgprb4tm4.1(flpo)And/J
029976 B6N.129S6-Retnlatm1Tliu/J
032924 B6N.129X1(B6J)-Msratm1Jmo/RlevMmjax
028764 B6N;129-Gt(ROSA)26Sortm1(CAG-GCaMP3)Dbe/J
030170 B6N;129-Gt(ROSA)26Sortm2(CAG-mGCaMP3)Dbe/J
007552 B6SJL-Chrdtm1Emdr/J
036061 BALB/c-Gt(ROSA)26Sortm1(CAG-EGFP,-BphP1/mCherry/TetR)Vvv/J
021568 BXSB.129(Cg)-Il21rtm1Wjl/DcrJ
017615 C.129(B6)-Trp53bp2tm1Cdlo/J
005700 C.129P2-Cxcr6tm1Litt/J
002724 C.129S2(B6)-Cxcr2tm1Mwm/J
017605 C.129S4-Icostm1Shr/J
002523 C.129S4-Myod1tm1Jae/J
002794 C.129S4-Tfap2atm1Jae/J
031371 C.129S6-Pglyrp3tm1Rdz/J
031372 C.129S6-Pglyrp4tm1Rdz/J
005417 C;129P2-Npy2rtm1Pern/J
003532 C3HeB.129S7-Kcna1tm1Tem/J
023520 C57BL/6-Bcrtm1(BCR/ABL)Tsr/J
031514 C57BL/6-Dnmt3atm1Rlvn/GrynvJ
035489 C57BL/6-Elmo1tm1Mki/J
038249 C57BL/6-Gt(ROSA)26Sortm1(CAG-Gria2*)Bli/J
029836 C57BL/6-Gt(ROSA)26Sortm1(CAG-SPAST*C448Y)Baas/J
035698 C57BL/6-Il17atm1Bcgen Il17fem1Litt/J
026685 C57BL/6-Nrxn1tm5Sud/J
017959 C57BL/6-Pmltm1(PML/RARA)Ley/J
038172 C57BL/6-Rgrtm1.1Kpal/J
038150 C57BL/6-Rxfp1tm2(RXFP1)Aia/J
035693 C57BL/6-Tlr9tm2Gbrt/J
038551 C57BL/6J-Gt(ROSA)26Sorem1(CAG-PACS1*R203W)Gath/J
038552 C57BL/6J-Gt(ROSA)26Sorem2(CAG-PACS1)Gath/J
035696 C57BL/6J-Saa3em1Litt/J
003219 D2.129P2(B6)-Nr5a1tm1Klp/EiJ
002787 DBA/1LacJ-Alox5aptm1Jdm/J
012440 DBA/2J-Tg(RP24-180N9)2Kjn/Kjn
026396 FVB.129S2(B6)-Bcrtm1Hkp/J
022036 FVB.129X1(Cg)-Atp2a2tm1Ges/Mmjax
013742 FVB.Cg-Slc9a1tm1Smb/Mmjax
027670 FVB/NJ-Tg(Snca)2Nbm/J
036631 FVB/NJ-Tg(Ube3a)5Mpan/J
004352 NOD.129S1(B6)-Ifngr2tm1Pbro/DvsJ
017637 NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ
030954 STOCK Acacatm1Jdh/J
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
033323 STOCK Adh7tm1Gdu/Mmjax
003810 STOCK Adrb1tm1Bkk Adrb2tm1Bkk/J
026229 STOCK Akap12tm1Ihg Rb1tm2Brn Tg(Pbsn-cre)4Prb/J
025665 STOCK Asxl1tm1.1Iaai/J
030647 STOCK Casrtm1Mrpk/J
006873 STOCK Cebpbtm1Vpo/J
026629 STOCK Cyp26b1tm1Dcp/J
026199 STOCK F8tm1.1Rmnt/J
017990 STOCK Fkbp4tm1Dvds/J
027233 STOCK Flnbtm1Vshn/J
038587 STOCK Foxi3em2(cre/ERT2)Akg/J
028096 STOCK Gata6tm1Hadj/J
038382 STOCK Ggta1tm1Jbl/NbndrJ
038583 STOCK Gt(ROSA)26Sorem1(CAG-Atoh1/EGFP)Akg/J
038584 STOCK Gt(ROSA)26Sorem2(CAG-Gf1i,-Atoh1/EGFP)Akg/J
038585 STOCK Gt(ROSA)26Sorem3(CAG-Gf1i,-Atoh1/EGFP,-Pou4f3)Akg/J
029812 STOCK Gt(ROSA)26Sortm1(RARA*)Soc/HsvJ
011004 STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm3(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J
037697 STOCK Gt(ROSA)26Sortm13(CAG-HRP)Luo/J
038177 STOCK Gt(ROSA)26Sortm2(CAG-APOBEC3B*E255A)Rshar/J
012266 STOCK Gt(ROSA)26Sortm5(ACTB-tTA)Luo/J
038629 STOCK Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze Pdgfrbtm1.1(flpo)Leol/LeolJ
023314 STOCK Homer3tm1Mhd/PfwJ
003259 STOCK Igf1tm2Ts/ImJ
022976 STOCK Igs2tm1(ACTB-EGFP,-tdTomato)Zng/J
003534 STOCK Inpp5dtm1Dmt/J
029787 STOCK Klf5tm1Jaw/J
023916 STOCK Lin28atm1.1Gqda/J
017508 STOCK Mir137tm1Mtm/Mmjax
013087 STOCK Mir7-2tm1Mtm/Mmjax
035925 STOCK Opn4tm1(cre)Saha/J
038627 STOCK Pdgfratm1(flpo)Leol Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze/LeolJ
038628 STOCK Pdgfratm2.1(flpo)Leol Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze/LeolJ
030357 STOCK Pou4f2tm1(cre)Bnt/J
022972 STOCK Pros1tm1Grl/J
027645 STOCK Rhotm1Jlem Tg(Rho*)2PRieke/J
027647 STOCK Rhotm1Jlem Tg(Rho*)STM2Rieke/J
027648 STOCK Rhotm1Jlem Tg(Rho*)TTM1Rieke/J
027649 STOCK Rhotm1Jlem Tg(Rho*)TTM4Rieke/J
037469 STOCK Sftpa1tm1Haw Tg(SFTPC-SFTPA1*)T1Jflo/Mmjax
036439 STOCK Slc2a2tm2.1Thor/J
012940 STOCK Snta1tm1Scf/J
030826 STOCK Srebf2tm1Jdh/J
010912 STOCK Wt1tm2(cre/ERT2)Wtp/J
035561 STOCK Tc(HSA21,CAG-EGFP)1Yakaz/J
025693 STOCK Tg(CAG-GFP,-mCherry/Pxn)#Mgin/KleyJ
007731 STOCK Tg(Col2a1-Npr2*B1)2Gar/J
005865 STOCK Tg(YACW408A5)1952Ricc/J
275 strains use this protocol