Protocol 22204: QPCR Assay - Generic Neo Quantitative PCR-QPCR- 1.2

B6(AU)-Tg(Map1a)56Pjn/Mmjax

Stock No: 036723

Version 8.0

Notes

Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler
480. Use an appropriate instrument specific Fluorophore/Quencher combination.
The transgene genotype is determined by comparing ΔCt values of each unknown
sample against known homozygous and hemizygous controls, using appropriate
endogenous references.

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

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
13741	Fluorophore-1	CAT CGC ATC GAG CGA GCA CGT ACT	Quencher-1	Tg Probe
oIMR1080		CGG CTG CAT ACG CTT GAT C		Transgene Forward	A
oIMR1081		CGA CAA GAC CGG CTT CCA T		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
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
oIMR1080	0.40 uM
oIMR1081	0.40 uM
oIMR1544	0.40 uM
oIMR3580	0.40 uM
Tg Probe	0.15 uM
IC Probe	0.15 uM
DNA

Cycling

Step	Temp °C	Time	Note
1	95.0	--
2	95.0	--
3	60.0	--	repeat steps 2-3 for 40 cycles

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
039976	129(Cg)-Gt(ROSA)26Sor^tm5(CAG-Sun1/sfGFP)Nat/CdnyJ
004166	129-Itgb5^tm1Des/J
037010	129;B6-Fmod^tm1Aol/SooJ
007664	129S-Efnb1^tm1Sor/J
008668	129S-Hba-a1^tm1Led/J
039968	129S-Slc6a4^tm2.1Rbl/JvvwJ
035806	129S4/SvJae-Fgfr2^tm6.1Sor/Mmjax
005032	B6 x B6Ei.129(Cg)-Bmp8b^tm1Blh/EiJ
036723	B6(AU)-Tg(Map1a)56Pjn/Mmjax
039527	B6(C)-Gt(ROSA)26Sor^em1.1(CAG-cas12a*)Sidi/J
039529	B6(C)-Gt(ROSA)26Sor^em2.1(CAG-cas12a*)Sidi/J
037318	B6(Cg)-H3c6^em1Sjes/J
028635	B6.129(Cg)-Cntnap2^tm2Pele/J
039525	B6.129(Cg)-Shank3^em1Yhj/J
037951	B6.129(Cg)-Wapl^tm1.1Kpfe/J
039307	B6.129-Clcn3^tm1Lamb/DnelJ
003558	B6.129-Grm5^tm1Rod/J
039728	B6.129-Gt(ROSA)26Sor^tm1(CAG-Sall4_ia,-Sall4_ib,-EGFP)Ykawa/J
037698	B6.129-Gt(ROSA)26Sor^tm13.1(CAG-HRP)Luo/J
030952	B6.129-Gt(ROSA)26Sor^tm5.1(CAG-Sun1/sfGFP)Nat/MmbeJ
038594	B6.129-H3f3a^tm1Hjf/J
034643	B6.129-Irgm1^tm1Gat/J
003870	B6.129-Plin1^tm1Chan/J
029902	B6.129-Ptprc^tm1Mak/Mmjax
034226	B6.129-Rab25^tm1Jrgo/J
003124	B6.129-Ucp1^tm1Kz/J
010725	B6.129-Ugt1^tm1Rhtu/J
017976	B6.129-Ulk1^tm1Thsn/J
036108	B6.129P2(C)-Cnr1^tm1.1Ltz/J
029253	B6.129P2(Cg)-Card6^tm1.1Aldu/MakJ
024630	B6.129P2-Lcn2^tm1Aade/AkiJ
034729	B6.129P2-Mylip^tm1Pton/J
005576	B6.129P2-P2rx7^tm1Gab/J
024839	B6.129P2-Pzp2^tm1Vln Mug1^tm2Vln/J
002122	B6.129P2-Tcrb^tm1Mom Tcrd^tm1Mom/J
023496	B6.129P3-Rasgrp3^tm1Jstn/TbwnJ
034595	B6.129S-Capn1^tm1Ahc/J
031661	B6.129S-Chat^tm1(cre)Lowl/MwarJ
040869	B6.129S-Eif2b5^tm1Oels/J
036668	B6.129S-Grpr^tm2(icre)Zfc/J
040456	B6.129S-Gt(ROSA)26Sor^tm1(CAG-Chrm3*,-mCitrine)Blr/Mmjax
038900	B6.129S-Mylk^tm2Avs/J
007015	B6.129S1-Gadd45g^tm1Flv/J
037386	B6.129S1-Gnat3^tm1Rfm/J
035475	B6.129S1-Ifi30^tm1Cres/J
038977	B6.129S1-Nfkb2^tm2Brv/AlexJ
004445	B6.129S2-Creb1^tm1Gsc/J
039731	B6.129S2-Dusp26^tm1Miv/J
040279	B6.129S2-Hsf4^tm1Miv/J
018311	B6.129S4-Catsper1^tm1Clph/J
039712	B6.129S4-Rel^tm1Hcl/PramJ
004185	B6.129S4-Soat2^tm1Far/J
037536	B6.129S4-Sypl2^tm1Hta/J
010672	B6.129S4-Tnfrsf11b^tm1Eac/J
038274	B6.129S4-Vps33b^tm1Zhen/J
039415	B6.129S6(Cg)-Asah1^tm1Medin/J
009340	B6.129S6(Cg)-Eif2ak3^tm1Dron/HotaJ
003742	B6.129S6-Cdh6^tm1Sma/J
017975	B6.129S6-Dag1^tm5Kcam/J
039210	B6.129S6-Tnfaip8l2^tm1Yhcn/J
002616	B6.129S7-Hba^tm1Paz/J
002277	B6.129S7-Src^tm1Sor/J
011131	B6.129X1(Cg)-Arrb1^tm1Jse/J
004225	B6.129X1-Zap70^tm1Weis/J
038957	B6.Cg-Cartpt^tm1(cre)Hze/J
035514	B6.Cg-Cd19^tm1(cre)Cgn Ikzf3^tm1.1Cjwu/J
037632	B6.Cg-Cd19^tm1.1(cre)Cgn Rps15^tm1.1Cjwu/J
036705	B6.Cg-Cd80^tm1Shr Cd86^tm2Shr Tg(Cd80)1Hode/J
039105	B6.Cg-Ces1g^tm1Reli/J
014162	B6.Cg-Fxn^tm1Mkn Fxn^tm1Pand/J
038190	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-Bcl2)Zhu/J
040470	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-Naaa)Dpio/J
039035	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-Tpp1)Plob/J
031279	B6.Cg-Gt(ROSA)26Sor^tm1(tetO-Yap1*,-GFP*)Stang/J
039236	B6.Cg-Gt(ROSA)26Sor^tm1.1(CAG-rtTA3,-GFP*,-luc*)Zhrt Tyr^c-2J/ZhrtJ
040817	B6.Cg-Gt(ROSA)26Sor^tm16(CAG-RRAS2,-EGFP)Geno/J
031842	B6.Cg-Irak1^tm1Jth/CpasJ
034191	B6.Cg-Mir21a^tm1Yoli/RthsnJ
038955	B6.Cg-Npr3^tm1(cre)Hze/J
040288	B6.Cg-Ptprc^a Pepc^b Tg(TcraTcrb)1100Mjb Gt(ROSA)26Sor^tm1.1(CAG-cas9*,-EGFP)Fezh/ShrJ
038958	B6.Cg-Rorb^tm1(cre)Hze/J
036415	B6.Cg-Sh2b1^em3Ccsu/J
038959	B6.Cg-Slc17a8^tm1(cre)Hze/J
034601	B6.Cg-Tgfb1^tm1Mki/J
038954	B6.Cg-Vipr2^em1(cre)Hze/J
033705	B6.Cg-Tg(Lck-RHO/ADRA1,-lacZ)B22-1Mik/J
021614	B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J
035020	B6.D1-Ksr2^tm1Rlw/J
036551	B6.FVB(Cg)-Tg(Pmch-cre)1Lowl/J
033109	B6;129-Cdh2^tm1(cre/ERT)Linli/J
038489	B6;129-Ces1d^tm2Reli/J
039578	B6;129-Eya1^tm1Px/J
039579	B6;129-Eya1^tm3(cre/ERT2)Px/J
039577	B6;129-Eya2^tm2Px/J
024649	B6;129-Gdf7^tm1Kng/J
036087	B6;129-Gt(ROSA)26Sor^tm3Cos/J
021039	B6;129-Gt(ROSA)26Sor^tm5(CAG-Sun1/sfGFP)Nat/J
039982	B6;129-Il1rl1^tm1Anjm/J
037191	B6;129-Nefh^tm2Dwc Nefm^tm1Mvr/J
006378	B6;129-Nrxn2^tm3Sud/J
018437	B6;129-Pax2^tm1Gdr/J
003255	B6;129-Plp1^tm1Kan/J
034559	B6;129-Prdm2^tm1Shg/J
038569	B6;129-Prop1^tm1Sac/Mmjax
036525	B6;129-Scgb1a1^tm1(icre)Fjd/J
039020	B6;129-Vgf^tm5Srjs/AbarJ
039865	B6;129P2-Nfkbie^tm1Bal/AlexJ
006374	B6;129P2-Rph3a^tm1Sud/J
002914	B6;129S-Hexb^tm1Rlp/J
003822	B6;129S-Psen1^tm1Shn/J
021968	B6;129S-Vcp^tm1Itl/J
035714	B6;129S1-Gt(ROSA)26Sor^tm1(Bcor*A)Vjba/J
003284	B6;129S1-Il1rap^tm1Roml/J
019578	B6;129S1-Rnls^tm1Gvd/J
002992	B6;129S1-Vcam1^tm1Roml/J
040387	B6;129S2-Gt(ROSA)26Sor^tm1(Gnas*R201C)Mrmn/J
036300	B6;129S4-Cux1^tm1(cre/ERT2)Zjh/J
028581	B6;129S4-Gt(ROSA)26Sor^tm2(CAG-HIST1H2BB/EGFP)Zjh/J
036304	B6;129S4-Gt(ROSA)26Sor^tm6(CAG-tTA)Zjh/J
039949	B6;129S4-Mir219a-2^tm1Qrlu Mir219a-1^tm1Qrlu/J
023343	B6;129S4-Pcmt1^tm1Scl/J
039106	B6;129S6-Ces1d^tm1Reli/J
023399	B6;129S6-Ctbp2^tm1Sud/J
034237	B6;129X-Stx2^tm1Dcru/J
039470	B6;129X1-Nfe2l1^tm1Jefc/J
006680	B6;CBA-Tg(Olfr16*,taulacZ)19Mom/MomJ
037014	B6J.129P2-Pofut2^Gt(RST434)Byg/BchMmjax
027410	B6J.129S1-Htt^tm1Mfc/190ChdiJ
370476	B6J.129S1-Htt^tm1Mfc/190JChdi
040162	B6JOlaHsd.129P2-Ercc1^tm1Jhjh/J
040164	B6JOlaHsd.129P2-Ercc1^tm2Jhjh/J
027971	B6N.129(Cg)-Pax6^tm1Pgr/J
025330	B6N.129P2(Cg)-Igs13^tm1Dolm Igs14^tm1Dolm/J
032924	B6N.129X1(B6J)-Msra^tm1Jmo/RlevMmjax
024744	B6N.Cg-Ids^tm1Muen/J
036061	BALB/c-Gt(ROSA)26Sor^tm1(CAG-EGFP,-BphP1/mCherry/TetR)Vvv/J
008340	BKS.Cg-Lepr^db Nos3^tm1Unc/RhrsJ
002724	C.129S2(B6)-Cxcr2^tm1Mwm/J
017605	C.129S4-Icos^tm1Shr/J
002523	C.129S4-Myod1^tm1Jae/J
031371	C.129S6-Pglyrp3^tm1Rdz/J
005417	C;129P2-Npy2r^tm1Pern/J
003532	C3HeB.129S7-Kcna1^tm1Tem/J
040918	C57BL/6-Ahr^tm1Lgzh/J
038023	C57BL/6-Cxcr3^tm2.1Arsa/J
035489	C57BL/6-Elmo1^tm1Mki/J
039526	C57BL/6-Gt(ROSA)26Sor^em1(CAG-cas12a*)Sidi/J
041116	C57BL/6-Gt(ROSA)26Sor^em1(CAG-KCNJ2)Tchn/J
038024	C57BL/6-Gt(ROSA)26Sor^em2(CAG-APPSwe/Ibe)Jjan/J
039528	C57BL/6-Gt(ROSA)26Sor^em2(CAG-cas12a*)Sidi/J
039508	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-CD63/copGFP)Yoy/J
038452	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Egfr*)Rnem/J
040844	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Got1)Chang/J
038249	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Gria2*)Bli/J
039534	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Plgrkt)Lmls/J
038454	C57BL/6-Gt(ROSA)26Sor^tm2(CAG-Egfr*L860R)Rnem/J
038535	C57BL/6-Gt(ROSA)26Sor^tm2(NGFR)Daav/J
035698	C57BL/6-Il17a^tm1Bcgen Il17f^em1Litt/J
026685	C57BL/6-Nrxn1^tm5Sud/J
017959	C57BL/6-Pml^tm1(PML/RARA)Ley/J
038172	C57BL/6-Rgr^tm1.1Kpal/J
038150	C57BL/6-Rxfp1^tm2(RXFP1)Aia/J
033221	C57BL/6-Sacs^tm2Bebr/J
035693	C57BL/6-Tlr9^tm2Gbrt/J
032131	C57BL/6-Tg(Scx-GFP/cre)1Stzr/J
038551	C57BL/6J-Gt(ROSA)26Sor^em1(CAG-PACS1*R203W)Gath/J
040505	C57BL/6J-Gt(ROSA)26Sor^em1(CAG-Sting1,-EGFP)Nyan/J
038552	C57BL/6J-Gt(ROSA)26Sor^em2(CAG-PACS1)Gath/J
035696	C57BL/6J-Saa3^em1Litt/J
007078	CByJ.129S2(B6)-Il6^tm1Kopf/J
002787	DBA/1LacJ-Alox5ap^tm1Jdm/J
040558	FVB.129S6-Gt(ROSA)26Sor^tm3(Pik3ca*E545K)Egan/J
040532	FVB.129S6-Gt(ROSA)26Sor^tm4(Pik3ca*E545K*H1047R)Egan/J
013742	FVB.Cg-Slc9a1^tm1Smb/Mmjax
036631	FVB/NJ-Tg(Ube3a)5Mpan/J
040163	FVBNHanHsd.129P2-Ercc1^tm1Jhjh/J
040165	FVBNHanHsd.129P2-Ercc1^tm2Jhjh/J
022760	MRL.Cg-Nos3^tm1Unc Fas^lpr/J
006023	NOD.Cg-H2-Ab1^b-tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ
039055	NOD.Cg-Prkdc^scid H2-K1^d-em1Dvs H2-Ab1^g7-em1Dvs H2-D1^b-em5Dvs Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/DvsJ
017637	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl H2-Ab1^b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ
040588	STOCK Actb^tm1Rsin/J
035185	STOCK Adgrl1^tm2c(EUCOMM)Hmgu/SudJ
003810	STOCK Adrb1^tm1Bkk Adrb2^tm1Bkk/J
030647	STOCK Casr^tm1Mrpk/J
006873	STOCK Cebpb^tm1Vpo/J
013701	STOCK Cep290^tm1Jgg/J
038587	STOCK Foxi3^em2(cre/ERT2)Akg/J
038382	STOCK Ggta1^tm1Jbl/NbndrJ
038583	STOCK Gt(ROSA)26Sor^em1(CAG-Atoh1/EGFP)Akg/J
038584	STOCK Gt(ROSA)26Sor^em2(CAG-Gf1i,-Atoh1/EGFP)Akg/J
038585	STOCK Gt(ROSA)26Sor^em3(CAG-Gf1i,-Atoh1/EGFP,-Pou4f3)Akg/J
038176	STOCK Gt(ROSA)26Sor^tm1(CAG-APOBEC3B)Rshar/J
039898	STOCK Gt(ROSA)26Sor^tm1(CAG-Atf4/EGFP,-tdTomato)Calak/J
038214	STOCK Gt(ROSA)26Sor^tm1(Stat6*)Voeh/J
037697	STOCK Gt(ROSA)26Sor^tm13(CAG-HRP)Luo/J
038177	STOCK Gt(ROSA)26Sor^tm2(CAG-APOBEC3B*E255A)Rshar/J
040009	STOCK Gt(ROSA)26Sor^tm3(tTA,tetO-TREM2,-EGFP)Gbu/NzhaoJ
040010	STOCK Gt(ROSA)26Sor^tm4(tTA,tetO-TREM2*R47H,-EGFP)Gbu/NzhaoJ
012266	STOCK Gt(ROSA)26Sor^tm5(ACTB-tTA)Luo/J
038629	STOCK Gt(ROSA)26Sor^tm65.2(CAG-tdTomato)Hze Pdgfrb^tm1.1(flpo)Leol/LeolJ
024390	STOCK Hat1^tm1Mrpa/J
003259	STOCK Igf1^tm2Ts/ImJ
039157	STOCK Lcn2^tm2Bara Tyr^c-2J/J
039204	STOCK Neurog2^tm1Cschu/J
030855	STOCK Notch1^tm2Rko Notch2^tm3Grid Gt(ROSA)26Sor^tm9(CAG-tdTomato)Hze Tg(KRT5-cre/ERT2)2Ipc/JeswJ
035925	STOCK Opn4^tm1(cre)Saha/J
038627	STOCK Pdgfra^tm1(flpo)Leol Gt(ROSA)26Sor^tm65.2(CAG-tdTomato)Hze/LeolJ
037681	STOCK Plk4^em4Ahol/J
038809	STOCK Proc^tm1Fjc Tg(F10-Proc)785Fjc/J
038808	STOCK Proc^tm1Fjc/J
027645	STOCK Rho^tm1Jlem Tg(Rho*)2PRieke/J
027647	STOCK Rho^tm1Jlem Tg(Rho*)STM2Rieke/J
027648	STOCK Rho^tm1Jlem Tg(Rho*)TTM1Rieke/J
027649	STOCK Rho^tm1Jlem Tg(Rho*)TTM4Rieke/J
037469	STOCK Sftpa1^tm1Haw Tg(SFTPC-SFTPA1*)T1Jflo/Mmjax
037902	STOCK Sftpc^tm1(dre/ERT2)Bzsh/J
036439	STOCK Slc2a2^tm2.1Thor/J
012940	STOCK Snta1^tm1Scf/J
039126	STOCK Spata7^tm1Mrd/RuchJ
033159	STOCK Utf1^tm1Jw/Mmjax
010912	STOCK Wt1^tm2(cre/ERT2)Wtp/J
035561	STOCK Tc(HSA21,CAG-EGFP)1Yakaz/J
223 strains use this protocol