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

B6.129X1(Cg)-Arrb1^tm1Jse/J

Stock No: 011131

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
ddH2O
Kapa Probe Fast QPCR	1.00 X
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
004166	129-Itgb5^tm1Des/J
037010	129;B6-Fmod^tm1Aol/SooJ
007664	129S-Efnb1^tm1Sor/J
008668	129S-Hba-a1^tm1Led/J
035806	129S4/SvJae-Fgfr2^tm6.1Sor/Mmjax
005032	B6 x B6Ei.129(Cg)-Bmp8b^tm1Blh/EiJ
037318	B6(Cg)-H3c6^em1Sjes/J
028635	B6.129(Cg)-Cntnap2^tm2Pele/J
037951	B6.129(Cg)-Wapl^tm1.1Kpfe/J
003558	B6.129-Grm5^tm1Rod/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
037191	B6.129-Nefh^tm2Dwc Nefm^tm1Mvr/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
024839	B6.129P2-Pzp^tm1Vln Mug1^tm2Vln/J
023496	B6.129P3-Rasgrp3^tm1Jstn/TbwnJ
034595	B6.129S-Capn1^tm1Ahc/J
031661	B6.129S-Chat^tm1(cre)Lowl/MwarJ
036668	B6.129S-Grpr^tm2(icre)Zfc/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
018311	B6.129S4-Catsper1^tm1Clph/J
004185	B6.129S4-Soat2^tm1Far/J
037536	B6.129S4-Sypl2^tm1Hta/J
010672	B6.129S4-Tnfrsf11b^tm1Eac/J
009340	B6.129S6(Cg)-Eif2ak3^tm1Dron/HotaJ
003742	B6.129S6-Cdh6^tm1Sma/J
017975	B6.129S6-Dag1^tm5Kcam/J
002277	B6.129S7-Src^tm1Sor/J
011131	B6.129X1(Cg)-Arrb1^tm1Jse/J
004225	B6.129X1-Zap70^tm1Weis/J
038214	B6.C(Cg)-Gt(ROSA)26Sor^tm1(Stat6*)Voeh/J
035514	B6.Cg-Cd19^tm1(cre)Cgn Ikzf3^tm1.1Cjwu/J
036705	B6.Cg-Cd80^tm1Shr Cd86^tm2Shr Tg(Cd80)1Hode/J
038176	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-APOBEC3B)Rshar/J
038190	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-Bcl2)Zhu/J
031279	B6.Cg-Gt(ROSA)26Sor^tm1(tetO-Yap1*,-GFP*)Stang/J
031842	B6.Cg-Irak1^tm1Jth/CpasJ
034191	B6.Cg-Mir21a^tm1Yoli/RthsnJ
036415	B6.Cg-Sh2b1^em3Ccsu/J
034601	B6.Cg-Tgfb1^tm1Mki/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
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
006378	B6;129-Nrxn2^tm3Sud/J
018437	B6;129-Pax2^tm1Gdr/J
003255	B6;129-Plp1^tm1Kan/J
034559	B6;129-Prdm2^tm1Shg/J
036525	B6;129-Scgb1a1^tm1(icre)Fjd/J
039020	B6;129-Vgf^tm5Srjs/AbarJ
006374	B6;129P2-Rph3a^tm1Sud/J
002914	B6;129S-Hexb^tm1Rlp/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
036304	B6;129S4-Gt(ROSA)26Sor^tm6(CAG-tTA)Zjh/J
023343	B6;129S4-Pcmt1^tm1Scl/J
034237	B6;129X-Stx2^tm1Dcru/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
034731	B6N(129S4)-Srd5a3^tm1c(EUCOMM)Wtsi/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
035489	C57BL/6-Elmo1^tm1Mki/J
038249	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Gria2*)Bli/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
035693	C57BL/6-Tlr9^tm2Gbrt/J
038551	C57BL/6J-Gt(ROSA)26Sor^em1(CAG-PACS1*R203W)Gath/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
013742	FVB.Cg-Slc9a1^tm1Smb/Mmjax
036631	FVB/NJ-Tg(Ube3a)5Mpan/J
022760	MRL.Cg-Nos3^tm1Unc Fas^lpr/J
006023	NOD.Cg-H2-Ab1^b-tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ
017637	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl H2-Ab1^b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ
035185	STOCK Adgrl1^tm2c(EUCOMM)Hmgu/SudJ
003810	STOCK Adrb1^tm1Bkk Adrb2^tm1Bkk/J
030647	STOCK Casr^tm1Mrpk/J
006873	STOCK Cebpb^tm1Vpo/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
037697	STOCK Gt(ROSA)26Sor^tm13(CAG-HRP)Luo/J
038177	STOCK Gt(ROSA)26Sor^tm2(CAG-APOBEC3B*E255A)Rshar/J
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
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
038628	STOCK Pdgfra^tm2.1(flpo)Leol Gt(ROSA)26Sor^tm65.2(CAG-tdTomato)Hze/LeolJ
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
036439	STOCK Slc2a2^tm2.1Thor/J
012940	STOCK Snta1^tm1Scf/J
033159	STOCK Utf1^tm1Jw/Mmjax
010912	STOCK Wt1^tm2(cre/ERT2)Wtp/J
035561	STOCK Tc(HSA21,CAG-EGFP)1Yakaz/J
025693	STOCK Tg(CAG-GFP,-mCherry/Pxn)#Mgin/KleyJ
143 strains use this protocol