Protocol 29498: Standard PCR Assay - Fluorescent Proteins (Generic GFP)

B6.Cg-Tg(Fos-tTA,Fos-EGFP*)1Mmay/J

Stock No: 018306

Version 4.2

Notes

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

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

Transgene = 173 bp
Internal positive control = 324 bp

Separated by gel electrophoresis on a 1.5% agarose gel.

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
oIMR0872		AAG TTC ATC TGC ACC ACC G		Transgene	A	GFP
oIMR1416		TCC TTG AAG AAG ATG GTG CG		Transgene	A	GFP
oIMR7338		CTA GGC CAC AGA ATT GAA AGA TCT		Internal Positive Control Forward	A
oIMR7339		GTA GGT GGA AAT TCT AGC ATC ATC C		Internal Positive Control Reverse	A

Reaction A

Component	Final Concentration
ddH2O
Kapa 2G HS buffer	1.30 X
MgCl2	2.60 mM
dNTP KAPA	0.26 mM
oIMR0872	0.50 uM
oIMR1416	0.50 uM
oIMR7338	0.50 uM
oIMR7339	0.50 uM
Glycerol	6.50 %
Dye	1.00 X
Kapa 2G HS taq polym	0.03 U/ul
DNA

Cycling

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

JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

JAX uses a 'touchdown' cycling protocol and therefore has not calculated the optimal annealing temperature for each set of primers.

Strains Using This Protocol

Stock Number	Strain Name
039976	129(Cg)-Gt(ROSA)26Sor^tm5(CAG-Sun1/sfGFP)Nat/CdnyJ
005483	129-Tg(CAG-EYFP)7AC5Nagy/J
034473	129;B6-Otx2^tm2(cre/ERT2)Tlam/J
032051	B6(129S4)-Kit^tm1.1(cre)Jmol/J
038534	B6(Cg)-Foxp3^tm1.1(flpo)Daav/J
039195	B6(Cg)-Il17a^tm1.1(EYFP/icre)Ehs/J
039963	B6(Cg)-Tyr^c-2J H2-Eb1^em1Jmlr/JmlrJ
038536	B6(Cg)-Tg(Cd8a-icre/ERT2,-GFP)#Daav/J
033943	B6.129(C)-Bdnf^tm1.1Bail/KmrtJ
035371	B6.129(Cg)-Msi2^tm1.1Geno/ReyaJ
004178	B6.129(Cg)-Tg(CAG-Bgeo/GFP)21Lbe/J
030420	B6.129(FVB)-Gt(ROSA)26Sor^tm1(CAG-birA,-EGFP)Joez/J
004218	B6.129(ICR)-Tg(CAG-ECFP)CK6Nagy/J
039728	B6.129-Gt(ROSA)26Sor^tm1(CAG-Sall4_ia,-Sall4_ib,-EGFP)Ykawa/J
030952	B6.129-Gt(ROSA)26Sor^tm5.1(CAG-Sun1/sfGFP)Nat/MmbeJ
032536	B6.129P2-Avil^tm2(cre)Fawa/J
036498	B6.Cg-Aire^tm2Mmat/EhrlJ
033942	B6.Cg-Bdnf^tm2.1Kmrt/J
039897	B6.Cg-Ccl3^tm1.1(HBEGF)Ckur/J
030305	B6.Cg-Eef1a1^tm1Rck/J
024953	B6.Cg-Gip^tm1.1Gmost/J
031279	B6.Cg-Gt(ROSA)26Sor^tm1(tetO-Yap1*,-GFP*)Stang/J
040817	B6.Cg-Gt(ROSA)26Sor^tm16(CAG-RRAS2,-EGFP)Geno/J
026944	B6.Cg-Gt(ROSA)26Sor^tm2.1(CAG-EGFP,-DTA*G128D)Pjen/J
037379	B6.Cg-Igs7^tm211(tetO-ChrimsonR/tdTomato,CAG-tTA2)Tasic/J
037380	B6.Cg-Igs7^tm214(CAG-ACR1*,CAG-mRuby3)Tasic/J
040737	B6.Cg-Ptprc^a Pepc^b Gt(ROSA)26Sor^tm1.1(CAG-cas9*,-EGFP)Fezh Tg(TcraTcrb)425Cbn/ShrJ
039726	B6.Cg-Sall4^tm1.1(EGFP/cre/ERT2)Ykawa/J
038835	B6.Cg-Sdk1^em1(cre/GFP)Jrs/KrishJ
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
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
018056	B6.FVB-Tg(CAG-boNT/B,-EGFP)U75-56Fwp/J
005738	B6.FVB-Tg(tetO-EGFP,-Tgfbr2)8Mcle/J
008126	B6.NOD-Tg(Cd4-EGFP)1Lt/J
021039	B6;129-Gt(ROSA)26Sor^tm5(CAG-Sun1/sfGFP)Nat/J
006665	B6;129P2-Or8a1^tm13(Olr226)Mom/MomJ
028581	B6;129S4-Gt(ROSA)26Sor^tm2(CAG-HIST1H2BB/EGFP)Zjh/J
038118	B6;129S6-Ank3^tm1Pmj/J
017495	B6;129S7-Crim1^tm1(GFP/cre/ERT2)Pzg/J
030537	B6;129S7-Drd2^tm1.1Jtw/J
034237	B6;129X-Stx2^tm1Dcru/J
039470	B6;129X1-Nfe2l1^tm1Jefc/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
031645	B6;D2-Tn(pb-CAG-dcas9,-EGFP)1Yangh/J
038774	B6;D2-Tg(PDGFB-SNCA/EGFP)78Ema/RoriJ
008344	B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J
025854	B6;FVB-Ptprc^a Tg(CAG-luc,-GFP)L2G85Chco Thy1^a/J
009159	B6;FVB-Tg(Cnp-EGFP/Rpl10a)JD368Htz/J
037194	B6;FVB-Tg(Cx3cl1-EGFP/Rpl10)ES26Htz/J
037198	B6;FVB-Tg(Gdnf-EGFP/Rpl10)ES2243Htz/J
004690	B6;FVB-Tg(Pcp2-EGFP)2Yuza/J
006043	B6;SJL-Tg(Oxt/EGFP)AI03Wsy/J
012350	B6;SJL-Tg(Thy1-COP4*H134R/EYFP)20Gfng/J
040426	B6J(B6N)-Tg(Nr4a1-EGFP/cre)820Khog/PalcaJ
004190	C.129-Il4^tm1Lky/J
040918	C57BL/6-Ahr^tm1Lgzh/J
038343	C57BL/6-Cnr2^tm1.1Geno/J
037626	C57BL/6-Gt(ROSA)26Sor^em1(CAG-Kcnq2*/EGFP)Hjch/J
039713	C57BL/6-Pgbd5^tm2.1Aken/J
039454	C57BL/6-Proser2^em3Odk/Mmjax
036868	C57BL/6-Rnf111^em1Litt/J
040504	C57BL/6-Sting1^em3Nyan/J
027662	C57BL/6-Tyr^c-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
005070	C57BL/6-Tg(Csf1r-EGFP-NGFR/FKBP1A/TNFRSF6)2Bck/J
017467	C57BL/6-Tg(Dmp1-Topaz)1Ikal/J
016617	C57BL/6-Tg(Nr4a1-EGFP/cre)820Khog/J
032131	C57BL/6-Tg(Scx-GFP/cre)1Stzr/J
007264	C57BL/6-Tg(Sry-EGFP)92Ei Tg(Sry)4Ei Chr Y^POS/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
040457	C57BL/6J-Gprc5a^em1(icre/ERT2)Blr/Mmjax
040505	C57BL/6J-Gt(ROSA)26Sor^em1(CAG-Sting1,-EGFP)Nyan/J
040416	C57BL/6J-Igs2^em1(CAG-mCherry,-Cxcr3*,-GFP*)Miji/J
040208	C57BL/6J-Igs2^em1(CAG-RFP*/GFP*)Qsun/J
036197	C57BL/6J-Nr1i2^em1Neg/J
041309	C57BL/6J-Tcf19^em1Jsun/J
038605	C57BL/6J-Tg(CAG-GFP*,-ATP6AP2)1Vli/J
006362	C57BL/6J-Tg(CMV-Cox8a/EYFP)17J/J
040140	C57BL/6J-Tg(Cxcl9-RFP,Cxcl10-BFP)1Adl/J
007857	C57BL/6J-Tg(Eno2-YFP/Cox8a)YRwb/J
007860	C57BL/6J-Tg(Eno2-YFP/Cox8a)ZRwb/J
007567	C57BL/6J-Tg(Itgax-cre,-EGFP)4097Ach/J
009593	C57BL/6J-Tg(Pomc-EGFP)1Low/J
039123	C57BL/6N-Trac^em1Gmrs/J
007075	CByJ.B6-Tg(CAG-EGFP)1Osb/J
007076	CByJ.B6-Tg(UBC-GFP)30Scha H2^b/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
028698	FVB/N-Tg(CAG-EGFP,-Tnf)1Kul/J
003257	FVB/N-Tg(GFAPGFP)14Mes/J
040440	NOD.B6(Cg)-Tg(Ins1-INS/sfGFP)369Arvn/J
013233	NOD.B6-Tg(Itgax-cre,-EGFP)4097Ach/J
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 Adgra3^tm1.1(HBEGF,-cre/ERT2)Pac/Mmjax
012881	STOCK Ascl1^tm1Reed/J
040665	STOCK Cntn2^tm1(DTA)Dkar/J
040090	STOCK Gli1^tm4.1Alj/J
006570	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Hlxb9-GFP)1Tmj/J
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
039898	STOCK Gt(ROSA)26Sor^tm1(CAG-Atf4/EGFP,-tdTomato)Calak/J
039717	STOCK Gt(ROSA)26Sor^tm16(CAG-Nsd2,-EBFP)Rsky/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
040818	STOCK Hoxa5^em1Dmwe/J
040717	STOCK Igs2^em1(HNF4A-Gsta2,-H2B/EGFP)Amc/J
034694	STOCK Igs7^tm86(tetO-CiVS/pHluorin)Hze/VpiJ
039204	STOCK Neurog2^tm1Cschu/J
039935	STOCK Piezo2^em1.1Ddg/J
039838	STOCK Pten^tm1.1Ayr/J
036906	STOCK Tmem178b^tm1c(KOMP)Wtsi Tmem178^tm1.1Sud/J
040657	STOCK Tmprss2^tm1.1(cre/ERT2)Ychen/J
039724	STOCK Vim^tm2(flpo)Ggen/J
010911	STOCK Wt1^tm1(EGFP/cre)Wtp/J
039156	STOCK Zic5^tm1.1(icre/ERT2)Akg/J
017472	STOCK Tg(Acp5-CFP,Ibsp-YFP,Dmp1-RFP)1Pmay/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
011106	STOCK Tg(CAG-GFP*)1Hadj/J
017468	STOCK Tg(Col1a1*3.6-Cyan)2Rowe/J
005854	STOCK Tg(Cp-EGFP)25Gaia/J
018322	STOCK Tg(Cp-EGFP)25Gaia/ReyaJ
039609	STOCK Tg(HBE1-H2B/CFP)30Maba/J
039607	STOCK Tg(HBE1-H2B/EGFP)55Maba/J
005418	STOCK Tg(HIST1H2BB/EGFP)1Pa/J
017952	STOCK Tg(Isl1-EGFP*)1Slp/J
039608	STOCK Tg(Kdr-H2B/EYFP)32Maba/J
012345	STOCK Tg(tetO-tdTomato,-Syp/EGFP*)1.1Luo/J
006129	STOCK Tg(Zp3-EGFP)1Dean/J
164 strains use this protocol