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Melting curve analysis is performed using a Roche Light Cycler 480.
PLEASE NOTE: This assay will distinguish heterozygous from homozygous Tcrd<tm1Mom> mutant mice.
Mutant Tm = 90°C +/- 1.0°C
Wild type Tm = 82°C+/- 1.0°C
Mut = 280 bp
IPC = 206 bp
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 |
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 |
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 |
Stock Number | Strain Name |
---|---|
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 |
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 |
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 |
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 |
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 |
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 |
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 |
038957 | B6.Cg-Cartpttm1(cre)Hze/J |
035514 | B6.Cg-Cd19tm1(cre)Cgn Ikzf3tm1.1Cjwu/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 |
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 |
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 |
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 |
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 |
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 |
016835 | B6;129S4-L3mbtl1tm1.1Haho/J |
003749 | B6;129S5-Pak2Gt(OST104058)Lex/J |
002594 | B6;129S6-Gba1tm1Nsb/J |
034237 | B6;129X-Stx2tm1Dcru/J |
027102 | B6;129X1-Lhcgrtm1Zmlei/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 |
034731 | B6N(129S4)-Srd5a3tm1c(EUCOMM)Wtsi/J |
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 |
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 |
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 |
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 |
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 |
029812 | STOCK Gt(ROSA)26Sortm1(RARA*)Soc/HsvJ |
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 |
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 |
037469 | STOCK Sftpa1tm1Haw Tg(SFTPC-SFTPA1*)T1Jflo/Mmjax |
036439 | STOCK Slc2a2tm2.1Thor/J |
012940 | STOCK Snta1tm1Scf/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 |
228 strains use this protocol |