Protocol 22106: Probe Assay - Gt(rosa)26sor<Tm1Sor> Probe

STOCK Hprt1^{tm65(Ple53-lacZ)Ems}/Mmjax

Stock No: 012353

Version 3.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. 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

Mut= 195 bp

Wt= 193 bp

Sequence

Wt Sequence:

 

CTGGCTTCTGAGGACCGCCCTGGGCCTGGGAGAATCCCTTCCCCCTCTTCCCTCGTGATCTGCAACTCCAGtctttctagaaGATGGGCGGG

AGTCTTCTGGGCAGGCTTAAAGGCTAACCTGGTGTGTGGGCGTTGTCCTGCAGGGGAATTGAACAGGTGTAAAATTGGAGGGACAAGACTTCCCACAGATT

Mutant Sequence:

CTGGCTTCTGAGGACCGCCCTGGGCCTGGGAGAATCCCTTCCCCCTCTTCCCTCGTGATCTGCAACTCCAGTCTTTCTAGtgGATCCCCCg

ggcTGCAGATctgTAGGGCGCAGTAGTCCAGGGTTTCCTTGATGATGTCATACTTATCCTGTCCCTTTTTTTTCCACAGCTCGCGGTTGAGGACAAACTCTTCG

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
21306		CTG GCT TCT GAG GAC CG		Common	A	Gt(ROSA)26Sor
21307		CGA AGA GTT TGT CCT CAA CCG		Mutant Reverse	A	SA site
21308	Fluorophore-1	ACC CTG GAC TAC TGC GCC C	Quencher-1	MUT Probe		SA site
21309		AAT CTG TGG GAA GTC TTG TCC		Wild type Reverse	A	Gt(ROSA)26Sor
21310	Fluorophore-2	TAA CCT GGT GTG TGG GCG TTG T	Quencher-2	WT Probe		Gt(ROSA)26Sor

Reaction A

Component	Final Concentration
Kapa Probe Fast QPCR	1.00 X
ddH2O
21306	0.40 uM
21307	0.40 uM
21309	0.40 uM
Wt Probe	0.15 uM
Mutant Probe	0.15 uM
DNA

Cycling

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

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
003310	129S-Gt(ROSA)26Sor^tm1Sor/J
007844	129S4/SvJae-Gt(ROSA)26Sor^tm2(FLP*)Sor/J
007689	129S4/SvJaeSor-Gt(ROSA)26Sor^tm4(attB/attP)Sor/J
009575	B6(129S4)-Et(cre/ERT2)119Rdav/J
012688	B6(129S4)-Et(cre/ERT2)13866Rdav/J
009582	B6(129S4)-Et(cre/ERT2)1645Rdav/J
009585	B6(129S4)-Et(cre/ERT2)2047Rdav/J
009574	B6(129S4)-Et(cre/ERT2)21Rdav/J
010688	B6(129S4)-Et(cre/ERT2)6691Rdav/J
010689	B6(129S4)-Et(cre/ERT2)6959Rdav/J
010690	B6(129S4)-Et(cre/ERT2)7089Rdav/J
010691	B6(129S4)-Et(cre/ERT2)7149Rdav/J
010692	B6(129S4)-Et(cre/ERT2)7381Rdav/J
010693	B6(129S4)-Et(cre/ERT2)8120Rdav/J
010694	B6(129S4)-Et(cre/ERT2)8131Rdav/J
010695	B6(129S4)-Et(cre/ERT2)9699Rdav/J
012690	B6(129S4)-Et(icre/ERT2)14208Rdav/J
012687	B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J
013730	B6(Cg)-Calb2^tm2.1(cre/ERT2)Zjh/J
012704	B6(Cg)-Crh^tm1(cre)Zjh/J
010705	B6(Cg)-Dlx5^tm1(cre/ERT2)Zjh/J
010776	B6(Cg)-Lhx6^tm1(cre/ERT2)Zjh/J
010777	B6(Cg)-Pvalb^tm1(cre/ERT2)Zjh/J
007708	B6.129-Gt(ROSA)26Sor^tm1(HD*103Q)Xwy/J
008877	B6.129P2(129S4)-Hprt1^tm12(Ple177-EGFP/cre)Ems/Mmjax
009114	B6.129P2(129S4)-Hprt1^tm14(Ple103-EGFP/cre)Ems/Mmjax
008709	B6.129P2(129S4)-Hprt1^tm9(Ple178-EGFP/cre)Ems/Mmjax
016933	B6.129P2(Cg)-Cdh1^tm1Cle/J
011008	B6.129P2(Cg)-Gt(ROSA)26Sor^tm1(tTA)Roos/J
009115	B6.129P2(Cg)-Hprt1^tm15(Ple111-EGFP)Ems/Mmjax
009348	B6.129P2(Cg)-Hprt1^tm17(Ple48-lacZ)Ems/Mmjax
009118	B6.129P2(Cg)-Hprt1^tm18(Ple90-EGFP)Ems/Mmjax
009596	B6.129P2(Cg)-Hprt1^tm33(Ple183-EGFP)Ems/Mmjax
010709	B6.129P2(Cg)-Hprt1^tm44(Ple49-lacZ)Ems/Mmjax
012580	B6.129P2(Cg)-Hprt1^tm59(Ple123-lacZ)Ems/Mmjax
012581	B6.129P2(Cg)-Hprt1^tm62(Ple153-lacZ)Ems/Mmjax
012583	B6.129P2(Cg)-Hprt1^tm68(Ple127-lacZ)Ems/Mmjax
008707	B6.129P2(Cg)-Hprt1^tm7(Ple185-EGFP)Ems/Mmjax
008708	B6.129P2(Cg)-Hprt1^tm8(Ple151-EGFP)Ems/Mmjax
034676	B6.129S(Cg)-Apoe^tm1(APOE*)Ljohn/J
003474	B6.129S4-Gt(ROSA)26Sor^tm1Sor/J
012930	B6.129S4-Gt(ROSA)26Sor^tm2(FLP*)Sor/J
007743	B6.129S4-Gt(ROSA)26Sor^tm3(phiC31*)Sor/J
010671	B6.129S4-Pkd1^tm2Ggg/J
009673	B6.129S6(C)-Gt(ROSA)26Sor^tm3(HIF1A*)Kael/J
032087	B6.129S6(Cg)-Gt(ROSA)26Sor^tm1(DTA)Jpmb/J
008466	B6.129X1(Cg)-Shh^tm6Amc/J
021071	B6.Cg-Gt(ROSA)26Sor^tm1(CAG-PA-GFP)Rmpl/J
026293	B6.Cg-Gt(ROSA)26Sor^tm1(LRRK2*R1441C)Djmo/J
029784	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm1(tetO-U2AF1*S34F)Mjwa/J
036937	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm2(tetO-CTNNB1*)Hoch Tg(Vav1-NUP98/HOXD13)G2Apla/MgkJ
014588	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm6(tetO-MSI2)Jae/J
017983	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm9(tetO-Dnmt3b_i1)Jae/J
014602	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm1(tetO-mCherry)Eggn/J
023749	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Tg(tetO-Pou5f1,-Sox2,-Klf4,-Myc)1Srn/J
006965	B6.Cg-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae/J
014536	B6.Cg-Hprt1^tm75(Ple143-lacZ)Ems/Mmjax
016868	B6.Cg-Sst^tm1.2(tTA2)Hze/J
021070	B6.Cg-Tg(Thy1-PA-GFP)6Rmpl/J
010527	B6;129-Gt(ROSA)26Sor^tm1(DTA)Mrc/J
016262	B6;129-Gt(ROSA)26Sor^tm1(Foxo1/GFP)Jke/J
017962	B6;129-Gt(ROSA)26Sor^tm1(RAC1*)Jkis/J
021025	B6;129-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm1(tetO-cre)Haho/J
006911	B6;129-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm2(tetO-Pou5f1)Jae/J
008883	B6;129-Gt(ROSA)26Sor^tm1(SNCA*A53T)Djmo/TmdJ
008889	B6;129-Gt(ROSA)26Sor^tm2(SNCA*119)Djmo/TmdJ
008886	B6;129-Gt(ROSA)26Sor^tm3(SNCA*E46K)Djmo/TmdJ
021039	B6;129-Gt(ROSA)26Sor^tm5(CAG-Sun1/sfGFP)Nat/J
023843	B6;129-Gt(ROSA)26Sor^tm6(Fzd6)Nat/J
026007	B6;129-Gt(ROSA)26Sor^tm8(cre/Esr1*)Nat/J
022516	B6;129S-Gt(ROSA)26Sor^tm1(Cdkn1c)Jfpa/J
012373	B6;129S-Hoxb1^tm1(cre)Og/J
014592	B6;129S4-Col1a1^tm1(tetO-mCherry)Eggn/J
011001	B6;129S4-Col1a1^tm1(tetO-Pou5f1,-Klf4,-Sox2,-Myc)Hoch/J
012464	B6;129S4-Foxd1^tm2(GFP/cre/ERT2)Amc/J
029415	B6;129S4-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm1(tetO-cas9)Sho/J
016836	B6;129S4-Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm7(tetO-HIST1H2BJ/GFP)Jae/J
003309	B6;129S4-Gt(ROSA)26Sor^tm1Sor/J
009576	B6;129S4-Et(cre/ERT2)278Rdav/J
017738	B6;SJL-Tg(Foxl1-cre)1Khk/J
019101	B6N.129S4(B6)-Gt(ROSA)26Sor^tm1Sor/CjDswJ
019013	B6N.129S6(Cg)-Gt(ROSA)26Sor^tm2(EGFP/cre)Alj/J
008603	C.129P2(B6)-Gt(ROSA)26Sor^tm1(tTA)Roos/J
002955	C.129S7-Gt(ROSA)26Sor/J
040844	C57BL/6-Gt(ROSA)26Sor^tm1(CAG-Got1)Chang/J
007900	C57BL/6-Gt(ROSA)26Sor^tm1(HBEGF)Awai/J
016097	C57BL/6-Tg(Car1-cre)5Flt/J
026863	D2.129S4(B6)-Gt(ROSA)26Sor^tm1Sor/SjJ
011034	FVB(Cg)-Tg(Ghrhr-cre)3242Lsk/J
006954	FVB-Tg(Ddx4-cre)1Dcas/J
009427	FVB.129S4(B6)-Gt(ROSA)26Sor^tm1Sor/J
016977	FVB.129S6-Gt(ROSA)26Sor^tm1(Pik3ca*H1047R)Egan/J
016603	NOD.B6-Gt(ROSA)26Sor^tm1(HBEGF)Awai/DvsJ
010910	STOCK Cort^tm1(cre)Zjh/J
010702	STOCK Gad2^tm1(cre/ERT2)Zjh/J
010802	STOCK Gad2^tm2(cre)Zjh/J
006331	STOCK Gt(ROSA)26Sor^tm1(DTA)Jpmb/J
022793	STOCK Gt(ROSA)26Sor^tm1(LRRK2*R1441C)Djmo/J
008159	STOCK Gt(ROSA)26Sor^tm1(Notch1)Dam/J
034693	STOCK Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Hprt1^tm1(tetO-KMT2A/MLLT3)Sguo/J
011004	STOCK Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm3(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J
011011	STOCK Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm4(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J
011013	STOCK Gt(ROSA)26Sor^tm1(rtTA*M2)Jae Col1a1^tm5(tetO-Pou5f1,-Klf4,-Myc)Jae/J
008600	STOCK Gt(ROSA)26Sor^tm1(tTA)Roos/J
025980	STOCK Gt(ROSA)26Sor^tm1.1(birA)Mejr Ep300^tm1.1Wtp/Mmjax
025983	STOCK Gt(ROSA)26Sor^tm1.1(birA)Mejr Mef2c^tm1.1Wtp/Mmjax
025978	STOCK Gt(ROSA)26Sor^tm1.1(birA)Mejr Nkx2-5^tm1.1Wtp/Mmjax
025977	STOCK Gt(ROSA)26Sor^tm1.1(birA)Mejr Zfpm2^tm1.1Wtp /Mmjax
018903	STOCK Gt(ROSA)26Sor^tm2(EGFP/cre)Alj/J
009674	STOCK Gt(ROSA)26Sor^tm4(HIF2A*)Kael/J
009349	STOCK Hprt1^tm31(Ple67-EGFP)Ems/Mmjax
009594	STOCK Hprt1^tm32(Ple112-EGFP)Ems/Mmjax
010707	STOCK Hprt1^tm37(lacZ)Ems/Mmjax
012335	STOCK Hprt1^tm50(Ple55-lacZ)Ems/Mmjax
012353	STOCK Hprt1^tm65(Ple53-lacZ)Ems/Mmjax
012584	STOCK Hprt1^tm69(Ple134-lacZ)Ems/Mmjax
016117	STOCK Nog^tm1.1Rmh/J
006953	STOCK Notch1^tm3(cre)Rko/J
028560	STOCK Snca^tm1.2Vlb Gt(ROSA)26Sor^tm1Sho/J
014581	STOCK Trpm8^tm1Apat/J
010908	STOCK Vip^tm1(cre)Zjh/J
010911	STOCK Wt1^tm1(EGFP/cre)Wtp/J
012691	STOCK Et(icre/ERT2)14374Rdav/J
012692	STOCK Et(icre/ERT2)14602Rdav/J
012693	STOCK Et(icre/ERT2)14624Rdav/J
007684	STOCK Tg(Atoh1-cre/Esr1*)14Fsh/J
008852	STOCK Tg(En2-cre)22Alj/J
023965	STOCK Tg(Krt17-EGFP)#Cou/J
008847	STOCK Tg(Msx2-cre/ERT)27Alj/J
014158	STOCK Tg(Pax4-cre)1Dam/J
009606	STOCK Tg(Six2-EGFP/cre)1Amc/J
131 strains use this protocol