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Melting curve analysis is done using a Roche Light Cycler 480.
This assay will NOT distinguish hemizygous from homozygous transgenic animals.
This protocol can also be used for YFP.
The Jackson Laboratory designed this protocol using GFP sequence, however, we have found that it can be used to genotype for other fluorescence proteins. It is possible that mismatches in the primer sequences may make data interpretation challenging for other fluorescence proteins.
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
10201 | AGT GCT TCA GCC GCT ACC | Transgene Forward | A | |||
10202 | GAA GAT GGT GCG CTC CTG | Transgene Reverse | A | |||
13731 | Fluorophore-1 | TTC AAG TCC GCC ATG CCC GAA | Quencher-1 | MUT Probe | ||
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 | |||
TmoIMR0105 | Fluorophore-2 | CCA ATG GTC GGG CAC TGC TCA A | Quencher-2 | IC Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
10201 | 0.40 uM |
10202 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Wt Probe | 0.15 uM |
Mutant Probe | 0.15 uM |
DNA |
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 |
Stock Number | Strain Name |
---|---|
005483 | 129-Tg(CAG-EYFP)7AC5Nagy/J |
034473 | 129;B6-Otx2tm2(cre/ERT2)Tlam/J |
016925 | 129;B6-Del(10Grin3b-Tmem259)1Zang Tg(Prnp-Tmem259,-EGFP)6Zhang/J |
021844 | 129S6.129P2(B6)-Gucy1b2tm2Mom/MomJ |
032051 | B6(129S4)-Kittm1.1(cre)Jmol/J |
033943 | B6.129(C)-Bdnftm1.1Bail/KmrtJ |
035371 | B6.129(Cg)-Msi2tm1.1Geno/ReyaJ |
029846 | B6.129(Cg)-Rettm13Jmi/SjnJ |
004178 | B6.129(Cg)-Tg(CAG-Bgeo/GFP)21Lbe/J |
018979 | B6.129(Cg)-Tg(CAG-EGFP)D4Nagy/KnwJ |
030420 | B6.129(FVB)-Gt(ROSA)26Sortm1(CAG-birA,-EGFP)Joez/J |
004218 | B6.129(ICR)-Tg(CAG-ECFP)CK6Nagy/J |
030952 | B6.129-Gt(ROSA)26Sortm5.1(CAG-Sun1/sfGFP)Nat/MmbeJ |
032536 | B6.129P2-Aviltm2(cre)Fawa/J |
028623 | B6.129P2-S1pr1tm1Hrose/J |
021930 | B6.129S1-Tg(CAG-EGFP)S1C2Tpo/KnwPeaJ |
031622 | B6.129S4-Ltb4r2tm1Bodd/J |
031621 | B6.129S4-Del(14Ltb4r2-Ltb4r1)1Bodd/J |
023046 | B6.129S6(Cg)-Ntrk2tm2.1Ddg/J |
012904 | B6.129S6-S100a4tm1Egn/YunkJ |
033942 | B6.Cg-Bdnftm2.1Kmrt/J |
030305 | B6.Cg-Eef1a1tm1Rck/J |
030541 | B6.Cg-Foxp2tm1.1(cre)Rpa/J |
024953 | B6.Cg-Giptm1.1Gmost/J |
031279 | B6.Cg-Gt(ROSA)26Sortm1(tetO-Yap1*,-GFP*)Stang/J |
026944 | B6.Cg-Gt(ROSA)26Sortm2.1(CAG-EGFP,-DTA*G128D)Pjen/J |
029043 | B6.Cg-Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze/J |
037379 | B6.Cg-Igs7tm211(tetO-ChrimsonR/tdTomato,CAG-tTA2)Tasic/J |
037380 | B6.Cg-Igs7tm214(CAG-ACR1*,CAG-mRuby3)Tasic/J |
030544 | B6.Cg-Npy1rtm1.1(cre/GFP)Rpa/J |
029236 | B6.Cg-Pabpn1tm1.1Gpvl/J |
022486 | B6.Cg-Ptprca Tg(UBC-PA-GFP)1Mnz/J |
028624 | B6.Cg-S1pr3tm1.1Hrose/J |
017466 | B6.Cg-Tg(Col1a1*3.6-Topaz)2Rowe/J |
030671 | B6.Cg-Tg(Col2a1-EGFP)1Waho/J |
018549 | B6.Cg-Tg(Csf1r-EGFP)1Hume/J |
018306 | B6.Cg-Tg(Fos-tTA,Fos-EGFP*)1Mmay/J |
014135 | B6.Cg-Tg(Fos/EGFP)1-3Brth/J |
007673 | B6.Cg-Tg(Gad1-EGFP)3Gfng/J |
006069 | B6.Cg-Tg(HIST1H2BB/EGFP)1Pa/J |
005029 | B6.Cg-Tg(Hlxb9-GFP)1Tmj/J |
006864 | B6.Cg-Tg(Ins1-EGFP)1Hara/J |
012643 | B6.Cg-Tg(Ly6a-EGFP)G5Dzk/J |
007894 | B6.Cg-Tg(Rgs4-EGFP)4Lvt/J |
022086 | B6.Cg-Tg(RP24-131B16/EGFP)13Ghan/J |
019494 | B6.Cg-Tg(RP24-131B16/EGFP)37Ghan/J |
007940 | B6.Cg-Tg(Thy1-CFP/COX8A)C1Lich/J |
007967 | B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J |
007615 | B6.Cg-Tg(Thy1-COP4/EYFP)9Gfng/J |
005630 | B6.Cg-Tg(Thy1-EYFP)15Jrs/J |
030257 | B6.FVB(Cg)-Tg(Colgalt2-EGFP/Rpl10a)DU9Htz/J |
018056 | B6.FVB-Tg(CAG-boNT/B,-EGFP)U75-56Fwp/J |
027496 | B6.FVB-Tg(Ckm-SLC2A4/EGFP)1Samc/J |
006000 | B6.FVB-Tg(ITGAM-HBEGF/EGFP)34Lan/J |
005738 | B6.FVB-Tg(tetO-EGFP,-Tgfbr2)8Mcle/J |
008126 | B6.NOD-Tg(Cd4-EGFP)1Lt/J |
021039 | B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J |
006664 | B6;129P2-Or6a2tm1Mom/MomJ |
006665 | B6;129P2-Or8a1tm13(Olr226)Mom/MomJ |
024750 | B6;129S4-Gt(ROSA)26Sortm9(EGFP/Rpl10a)Amc/J |
017495 | B6;129S7-Crim1tm1(GFP/cre/ERT2)Pzg/J |
030537 | B6;129S7-Drd2tm1.1Jtw/J |
019908 | B6;129S7-Ghsrtm2Rgs/J |
034237 | B6;129X-Stx2tm1Dcru/J |
025820 | B6;CBA-Tg(CRH-MAPT/Topaz)1Rck/J |
033639 | B6;CBA-Tg(Gnrh1-EGFP)51Sumo/J |
007986 | B6;CBA-Tg(H*/Olfr16-GFP)11Mom/MomJ |
007987 | B6;CBA-Tg(H*/Olfr16-GFP)25Mom/MomJ |
007979 | B6;CBA-Tg(H/Olfr16-GFP)3Mom/MomJ |
007980 | B6;CBA-Tg(H/Olfr16-GFP)4Mom/MomJ |
007984 | B6;CBA-Tg(H/Olfr16-taumCherry,-tauGFP)11Mom/MomJ |
007985 | B6;CBA-Tg(H/Olfr16-taumCherry,-tauGFP)13Mom/MomJ |
007977 | B6;CBA-Tg(Hf/Olfr16-GFP)7Mom/MomJ |
006614 | B6;CBA-Tg(Thy1-CFP/COX8A)C1Lich/J |
006617 | B6;CBA-Tg(Thy1-CFP/COX8A)S2Lich/J |
038774 | B6;D2-Tg(PDGFB-SNCA/EGFP)78Ema/RoriJ |
005621 | B6;D2-Tg(S100B-EGFP)1Wjt/J |
031645 | B6;D2-Tn(pb-CAG-cas9*,-EGFP)1Yangh/J |
008344 | B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J |
025854 | B6;FVB-Ptprca Tg(CAG-luc,-GFP)L2G85Chco Thy1a/J |
009159 | B6;FVB-Tg(Cnp-EGFP/Rpl10a)JD368Htz/J |
030252 | B6;FVB-Tg(Cort-EGFP/Rpl10a)GM130Htz/J |
037194 | B6;FVB-Tg(Cx3cl1-EGFP/Rpl10)ES26Htz/J |
030255 | B6;FVB-Tg(Drd2-EGFP/Rpl10a)CP101Htz/J |
037198 | B6;FVB-Tg(Gdnf-EGFP/Rpl10)ES2243Htz/J |
030265 | B6;FVB-Tg(Olig2-EGFP/Rpl10a)JD97Htz/J |
004690 | B6;FVB-Tg(Pcp2-EGFP)2Yuza/J |
030271 | B6;FVB-Tg(Sept4-EGFP/Rpl10a)DS152Htz/J |
006043 | B6;SJL-Tg(Oxt/EGFP)AI03Wsy/J |
012350 | B6;SJL-Tg(Thy1-COP4*H134R/EYFP)20Gfng/J |
035474 | B6N.Cg-Tg(Arc-tTA,-EGFP/Capzb*)10Mkyph/J |
004190 | C.129-Il4tm1Lky/J |
005700 | C.129P2-Cxcr6tm1Litt/J |
023520 | C57BL/6-Bcrtm1(BCR/ABL)Tsr/J |
038343 | C57BL/6-Cnr2tm1.1Geno/J |
037626 | C57BL/6-Gt(ROSA)26Sorem1(CAG-Kcnq2*/EGFP)Hjch/J |
029316 | C57BL/6-Gt(ROSA)26Sortm1(CAG-FOXO1,GFP)Moli/J |
036868 | C57BL/6-Rnf111em1Litt/J |
027662 | C57BL/6-Tyrc-Brd Tg(Gh1-luc/EGFP)D8Mrln/J |
017469 | C57BL/6-Tg(BGLAP-Topaz)1Rowe/J |
006567 | C57BL/6-Tg(CAG-EGFP)131Osb/LeySopJ |
003291 | C57BL/6-Tg(CAG-EGFP)1Osb/J |
029889 | C57BL/6-Tg(CAG-tdTomato,Actb/EGFP)1Erv/J |
028596 | C57BL/6-Tg(Cck-EGFP)2Mirn/J |
005070 | C57BL/6-Tg(Csf1r-EGFP-NGFR/FKBP1A/TNFRSF6)2Bck/J |
017467 | C57BL/6-Tg(Dmp1-Topaz)1Ikal/J |
007264 | C57BL/6-Tg(Sry-EGFP)92Ei Tg(Sry)4Ei Chr YPOS/EiJ |
005706 | C57BL/6-Tg(tetO-CDK5R1/GFP)337Lht/J |
010713 | C57BL/6-Tg(tetO-GFP/tetX)5696Stl/J |
004353 | C57BL/6-Tg(UBC-GFP)30Scha/J |
036197 | C57BL/6J-Nr1i2em1Neg/J |
027898 | C57BL/6J-Tg(CAG-lacZ,-FUS,-EGFP)629Gyu/J |
007857 | C57BL/6J-Tg(Eno2-YFP/Cox8a)YRwb/J |
007860 | C57BL/6J-Tg(Eno2-YFP/Cox8a)ZRwb/J |
009593 | C57BL/6J-Tg(Pomc-EGFP)1Low/J |
028324 | C57BL/6NJ-Gpr15em2J/J |
007677 | CB6-Tg(Gad1-EGFP)G42Zjh/J |
007075 | CByJ.B6-Tg(CAG-EGFP)1Osb/J |
007076 | CByJ.B6-Tg(UBC-GFP)30Scha H2b/J |
026855 | D2.Cg-Tg(Thy1-Brainbow1.1)MLich/SjJ |
025019 | D2.Cg-Tg(Thy1-YFP/Syp)10Jrs/SjJ |
026856 | D2.FVB-Tg(GFAPGFP)14Mes/SjJ |
021187 | FVB(B6)-Tg(Pbsn-rtTA*M2)42Xy/J |
003718 | FVB-Tg(GadGFP)45704Swn/J |
005515 | FVB-Tg(ITGAM-HBEGF/EGFP)34Lan/J |
017484 | FVB-Tg(JPH3-GFP,-JPH3*)GXwy/J |
018071 | FVB-Tg(Prism)1861Htz/J |
003516 | FVB.Cg-Tg(CAG-EGFP)B5Nagy/J |
033064 | FVB/N-Tg(ACTA1-ATP2A1*/DsRed/GFP)5Wheel/J |
028698 | FVB/N-Tg(CAG-EGFP,-Tnf)1Kul/J |
003257 | FVB/N-Tg(GFAPGFP)14Mes/J |
027665 | FVB/N-Tg(Gnrhr-luc/EGFP)L8Mrln/J |
021937 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-EGFP)1Osb/SzJ |
008173 | NOD.Cg-Tg(Ins1-EGFP)1Hara/QtngJ |
009422 | NOD.Cg-Tg(Itgax-Venus)1Mnz/QtngJ |
005076 | NOD.Cg-Tg(tetO-EGFP/FADD)1Doi/DoiJ |
010542 | NOD.FVB-Tg(CAG-luc,-GFP)L2G85Chco/FathJ |
008547 | NOD.FVB-Tg(ITGAM-HBEGF/EGFP)34Lan/JdkJ |
008694 | NOD/ShiLt-Tg(Foxp3-EGFP/cre)1cJbs/J |
005282 | NOD/ShiLtJ-Tg(Ins1-EGFP/GH1)14Hara/HaraJ |
036735 | STOCK Adgra3tm1.1(HBEGF,-cre/ERT2)Pac/Mmjax |
027012 | STOCK Foxj1tm1.1(cre/ERT2/GFP)Htg/J |
032003 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Hlxb9-GFP)1Tmj Tg(SMN2*delta7)4299Ahmb/J |
006570 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Hlxb9-GFP)1Tmj/J |
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 |
024284 | STOCK Hcn4tm3Sev/J |
034694 | STOCK Igs7tm86(tetO-CiVS/pHluorin)Hze/VpiJ |
023449 | STOCK Myh14tm3.1Rsad/Mmjax |
025855 | STOCK Ptprca Lag3tm1Doi Tg(CAG-luc,-GFP)L2G85Chco Thy1a/J |
036906 | STOCK Tmem178btm1c(KOMP)Wtsi Tmem178tm1.1Sud/J |
010911 | STOCK Wt1tm1(EGFP/cre)Wtp/J |
017472 | STOCK Tg(Acp5-CFP,Ibsp-YFP,Dmp1-RFP)1Pmay/J |
026033 | STOCK Tg(Aldh1l1-EGFP,-DTA)D8Rth/J |
006850 | STOCK Tg(CAG-Bgeo,-NOTCH1,-EGFP)1Lbe/J |
003920 | STOCK Tg(CAG-Bgeo/GFP)21Lbe/J |
003115 | STOCK Tg(CAG-EGFP)B5Nagy/J |
003116 | STOCK Tg(CAG-EGFP)D4Nagy/J |
017919 | STOCK Tg(CAG-EGFP,-dsRed2/RNAi:Tardbp)6Zxu/J |
011106 | STOCK Tg(CAG-GFP*)1Hadj/J |
025693 | STOCK Tg(CAG-GFP,-mCherry/Pxn)#Mgin/KleyJ |
017468 | STOCK Tg(Col1a1*3.6-Cyan)2Rowe/J |
005854 | STOCK Tg(Cp-EGFP)25Gaia/J |
018322 | STOCK Tg(Cp-EGFP)25Gaia/ReyaJ |
026104 | STOCK Tg(Egfl7-EGFP)12Stmn/J |
005418 | STOCK Tg(HIST1H2BB/EGFP)1Pa/J |
017952 | STOCK Tg(Isl1-EGFP*)1Slp/J |
018148 | STOCK Tg(Slc17a8-EGFP)1Edw/SealJ |
012345 | STOCK Tg(tetO-tdTomato,-Syp/EGFP*)1.1Luo/J |
025195 | STOCK Tg(Vmn1r191-Mapt/CFP)1Dlc/J |
025194 | STOCK Tg(Vmn1r206-Mapt/YFP)1Dlc/J |
025193 | STOCK Tg(Vmn1r232-Mapt/EGFP)1Dlc/J |
006129 | STOCK Tg(Zp3-EGFP)1Dean/J |
174 strains use this protocol |