Stock No: 029904
Protocol 29086: Standard PCR Assay - Generic Neo
Version 2.3

Notes

Melting curve analysis is performed using a Roche Light Cycler 480.

PLEASE NOTE: This assay will distinguish heterozygous from homozygous Tcrd<tm1Mom> mutant mice.

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 Tm = 90°C +/- 1.0°C
Wild type Tm = 82°C+/- 1.0°C

Mut = 280 bp

IPC = 206 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR6916 CTT GGG TGG AGA GGC TAT TC Mutant Forward A Neo
oIMR6917 AGG TGA GAT GAC AGG AGA TC Mutant Reverse A Neo
oIMR8744 CAA ATG TTG CTT GTC TGG TG Internal Positive Control Forward A
oIMR8745 GTC AGT CGA GTG CAC AGT TT Internal Positive Control Reverse A

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
oIMR6916 0.50 uM
oIMR6917 0.50 uM
oIMR8744 0.50 uM
oIMR8745 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
039976 129(Cg)-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/CdnyJ
025451 129-Iqgap1tm1Aber/VsJ
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
007005 129S-Scg5tm1Led/J
018424 129S-Slc26a4tm1Egr/AjgJ
039968 129S-Slc6a4tm2.1Rbl/JvvwJ
035806 129S4/SvJae-Fgfr2tm6.1Sor/Mmjax
016264 129S6-Fmn2tm1Led/RwwJ
005032 B6 x B6Ei.129(Cg)-Bmp8btm1Blh/EiJ
036723 B6(AU)-Tg(Map1a)56Pjn/Mmjax
039527 B6(C)-Gt(ROSA)26Sorem1.1(CAG-cas12a*)Sidi/J
039529 B6(C)-Gt(ROSA)26Sorem2.1(CAG-cas12a*)Sidi/J
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
023313 B6.129(Cg)-Homer2tm1Mhd/PfwJ
008320 B6.129(Cg)-Leprtm2(cre)Rck/J
039525 B6.129(Cg)-Shank3em1Yhj/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
039728 B6.129-Gt(ROSA)26Sortm1(CAG-Sall4_ia,-Sall4_ib,-EGFP)Ykawa/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
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
002725 B6.129-Ptprctm1Holm/J
034226 B6.129-Rab25tm1Jrgo/J
006833 B6.129-Sgcbtm1Kcam/2J
023309 B6.129-Stat2tm1Shnd/J
036108 B6.129P2(C)-Cnr1tm1.1Ltz/J
022364 B6.129P2(SJL)-Ntrk3tm1Ddg/J
022499 B6.129P2-Adora2btm1Till/J
023726 B6.129P2-C1qatm1.1Tenn/J
029905 B6.129P2-Ctla4tm1Mak/Mmjax
005814 B6.129P2-Grm1tm1Dgen/J
002204 B6.129P2-Hbbtm1Unc/J
029904 B6.129P2-Msh2tm1Mak/Mmjax
034729 B6.129P2-Myliptm1Pton/J
002122 B6.129P2-Tcrbtm1Mom Tcrdtm1Mom/J
002120 B6.129P2-Tcrdtm1Mom/J
005845 B6.129P2-Thbs4tm1Dgen/J
034595 B6.129S-Capn1tm1Ahc/J
036668 B6.129S-Grprtm2(icre)Zfc/J
027187 B6.129S-Lnpeptm1Ske/J
038900 B6.129S-Mylktm2Avs/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
004445 B6.129S2-Creb1tm1Gsc/J
039731 B6.129S2-Dusp26tm1Miv/J
040279 B6.129S2-Hsf4tm1Miv/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
039712 B6.129S4-Reltm1Hcl/PramJ
004185 B6.129S4-Soat2tm1Far/J
037536 B6.129S4-Sypl2tm1Hta/J
010672 B6.129S4-Tnfrsf11btm1Eac/J
039415 B6.129S6(Cg)-Asah1tm1Medin/J
018648 B6.129S6(Cg)-Itgb2tm1.1Mskim/J
003742 B6.129S6-Cdh6tm1Sma/J
017975 B6.129S6-Dag1tm5Kcam/J
018164 B6.129S6-Pdcd4tm1Yhcn/J
039210 B6.129S6-Tnfaip8l2tm1Yhcn/J
025929 B6.129S7(Cg)-Pik3catm1Jdo/J
002616 B6.129S7-Hbatm1Paz/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
038957 B6.Cg-Cartpttm1(cre)Hze/J
035514 B6.Cg-Cd19tm1(cre)Cgn Ikzf3tm1.1Cjwu/J
039105 B6.Cg-Ces1gtm1Reli/J
014162 B6.Cg-Fxntm1Mkn Fxntm1Pand/J
038190 B6.Cg-Gt(ROSA)26Sortm1(CAG-Bcl2)Zhu/J
030743 B6.Cg-Gt(ROSA)26Sortm1(CAG-Pycard/mCitrine*,-CD2*)Dtg/J
039035 B6.Cg-Gt(ROSA)26Sortm1(CAG-Tpp1)Plob/J
040125 B6.Cg-Gt(ROSA)26Sortm1(Sirt6)Mpgu/J
031279 B6.Cg-Gt(ROSA)26Sortm1(tetO-Yap1*,-GFP*)Stang/J
039236 B6.Cg-Gt(ROSA)26Sortm1.1(CAG-rtTA3,-GFP*,-luc*)Zhrt Tyrc-2J/ZhrtJ
040009 B6.Cg-Gt(ROSA)26Sortm3(tTA,tetO-TREM2,-EGFP)Gbu/NzhaoJ
040010 B6.Cg-Gt(ROSA)26Sortm4(tTA,tetO-TREM2*R47H,-EGFP)Gbu/NzhaoJ
006513 B6.Cg-Ighmbp2nmd-2J Tg(Ttn-Ighmbp2)45Cx/Cx
039157 B6.Cg-Lcn2tm2Bara Tyrc-2J/J
024469 B6.Cg-Lrrk2tm3.1Mjfa/J
034191 B6.Cg-Mir21atm1Yoli/RthsnJ
027748 B6.Cg-Mybl1tm1Epr/J
038955 B6.Cg-Npr3tm1(cre)Hze/J
029031 B6.Cg-Pabpc1ltm1.1Esel/J
038958 B6.Cg-Rorbtm1(cre)Hze/J
036415 B6.Cg-Sh2b1em3Ccsu/J
038959 B6.Cg-Slc17a8tm1(cre)Hze/J
013717 B6.Cg-Slc20a1tm1.2Cmg/J
012627 B6.Cg-Stard4tm1.1Bres/J
034601 B6.Cg-Tgfb1tm1Mki/J
038954 B6.Cg-Vipr2em1(cre)Hze/J
033705 B6.Cg-Tg(Lck-RHO/ADRA1,-lacZ)B22-1Mik/J
008430 B6.Cg-Tg(Tcrb)HRBAll/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
039578 B6;129-Eya1tm1Px/J
039579 B6;129-Eya1tm3(cre/ERT2)Px/J
039577 B6;129-Eya2tm2Px/J
012820 B6;129-Fzd1tm1.1Nat/J
010948 B6;129-Gabra1tm4Geh/J
036087 B6;129-Gt(ROSA)26Sortm3Cos/J
021039 B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J
039982 B6;129-Il1rl1tm1Anjm/J
012894 B6;129-Macf1tm1Efu/J
006904 B6;129-Msctm1Eno/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
039020 B6;129-Vgftm5Srjs/AbarJ
002121 B6;129P-Tcrbtm1Mom Tcrdtm1Mom/J
039865 B6;129P2-Nfkbietm1Bal/AlexJ
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
040387 B6;129S2-Gt(ROSA)26Sortm1(Gnas*R201C)Mrmn/J
029304 B6;129S2-Syngap1tm2Geno/RumbJ
012623 B6;129S4-Carftm1.1Aew/J
036300 B6;129S4-Cux1tm1(cre/ERT2)Zjh/J
028581 B6;129S4-Gt(ROSA)26Sortm2(CAG-HIST1H2BB/EGFP)Zjh/J
016835 B6;129S4-L3mbtl1tm1.1Haho/J
003749 B6;129S5-Pak2Gt(OST104058)Lex/J
039106 B6;129S6-Ces1dtm1Reli/J
023399 B6;129S6-Ctbp2tm1Sud/J
002594 B6;129S6-Gba1tm1Nsb/J
034237 B6;129X-Stx2tm1Dcru/J
027102 B6;129X1-Lhcgrtm1Zmlei/J
039470 B6;129X1-Nfe2l1tm1Jefc/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
030975 B6J.129S7-Pamtm1Jep/Mmjax
027971 B6N.129(Cg)-Pax6tm1Pgr/J
021078 B6N.129S1-Mrgprb4tm4.1(flpo)And/J
029976 B6N.129S6-Retnlatm1Tliu/J
032924 B6N.129X1(B6J)-Msratm1Jmo/RlevMmjax
007552 B6SJL-Chrdtm1Emdr/J
021568 BXSB.129(Cg)-Il21rtm1Wjl/DcrJ
017615 C.129(B6)-Trp53bp2tm1Cdlo/J
005700 C.129P2-Cxcr6tm1Litt/J
017605 C.129S4-Icostm1Shr/J
002794 C.129S4-Tfap2atm1Jae/J
031371 C.129S6-Pglyrp3tm1Rdz/J
005417 C;129P2-Npy2rtm1Pern/J
003532 C3HeB.129S7-Kcna1tm1Tem/J
023520 C57BL/6-Bcrtm1(BCR/ABL)Tsr/J
038023 C57BL/6-Cxcr3tm2.1Arsa/J
035489 C57BL/6-Elmo1tm1Mki/J
039526 C57BL/6-Gt(ROSA)26Sorem1(CAG-cas12a*)Sidi/J
038024 C57BL/6-Gt(ROSA)26Sorem2(CAG-APPSwe/Ibe)Jjan/J
039528 C57BL/6-Gt(ROSA)26Sorem2(CAG-cas12a*)Sidi/J
039508 C57BL/6-Gt(ROSA)26Sortm1(CAG-CD63/copGFP)Yoy/J
038452 C57BL/6-Gt(ROSA)26Sortm1(CAG-Egfr*)Rnem/J
038249 C57BL/6-Gt(ROSA)26Sortm1(CAG-Gria2*)Bli/J
039534 C57BL/6-Gt(ROSA)26Sortm1(CAG-Plgrkt)Lmls/J
029836 C57BL/6-Gt(ROSA)26Sortm1(CAG-SPAST*C448Y)Baas/J
038454 C57BL/6-Gt(ROSA)26Sortm2(CAG-Egfr*L860R)Rnem/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
035693 C57BL/6-Tlr9tm2Gbrt/J
032131 C57BL/6-Tg(Scx-GFP/cre)1Stzr/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
009645 CB6-Isl1tm1Tmj/J
008312 CBy.129P2(B6)-Tcrdtm1Mom/SzJ
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
022395 NOD.129P2(B6)-Tcrdtm1Mom/JsdJ
004352 NOD.129S1(B6)-Ifngr2tm1Pbro/DvsJ
039055 NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/DvsJ
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
026229 STOCK Akap12tm1Ihg Rb1tm2Brn Tg(Pbsn-cre)4Prb/J
025665 STOCK Asxl1tm1.1Iaai/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
038176 STOCK Gt(ROSA)26Sortm1(CAG-APOBEC3B)Rshar/J
029812 STOCK Gt(ROSA)26Sortm1(RARA*)Soc/HsvJ
038214 STOCK Gt(ROSA)26Sortm1(Stat6*)Voeh/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
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
039204 STOCK Neurog2tm1Cschu/J
035925 STOCK Opn4tm1(cre)Saha/J
038627 STOCK Pdgfratm1(flpo)Leol Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze/LeolJ
037681 STOCK Plk4em4Ahol/J
030357 STOCK Pou4f2tm1(cre)Bnt/J
038809 STOCK Proctm1Fjc Tg(F10-Proc)785Fjc/J
038808 STOCK Proctm1Fjc/J
022972 STOCK Pros1tm1Grl/J
037469 STOCK Sftpa1tm1Haw Tg(SFTPC-SFTPA1*)T1Jflo/Mmjax
037902 STOCK Sftpctm1(dre/ERT2)Bzsh/J
036439 STOCK Slc2a2tm2.1Thor/J
012940 STOCK Snta1tm1Scf/J
039126 STOCK Spata7tm1Mrd/RuchJ
035561 STOCK Tc(HSA21,CAG-EGFP)1Yakaz/J
007731 STOCK Tg(Col2a1-Npr2*B1)2Gar/J
005865 STOCK Tg(YACW408A5)1952Ricc/J
274 strains use this protocol