Stock Number: 008549 Strain Name: NOD.FVB-Tg(Itgax-DTR/EGFP)57Lan/JdkJ

Allele: Generic GFP Protocol Name: Fluorescent Proteins (Generic GFP) Method: Standard PCR Version: 3.0 Created: 17-January -2011 Updated: 05-June -2011

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 173 bp Internal positive control = 324 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

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

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 5 U/ul KAPATaq DNA Polymerase 0.12 0.05 U/ul 0.12 ddH2O 3.30 - 3.30 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR0872 0.60 1.00 uM 0.60 20 uM oIMR1416 0.60 1.00 uM 0.60 20 uM oIMR7338 0.30 0.50 uM 0.30 20 uM oIMR7339 0.30 0.50 uM 0.30 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 DNA 2.00 - 2.00 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20

Cycling  Step # Temp °C Time Note 1 94 1.5 min - 2 94 30 sec - 3 60 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 003115 STOCK Tg(CAG-EGFP)B5Nagy/J 003116 STOCK Tg(CAG-EGFP)D4Nagy/J 003257 FVB/N-Tg(GFAPGFP)14Mes/J 003291 C57BL/6-Tg(CAG-EGFP)1Osb/J 003516 FVB.Cg-Tg(CAG-EGFP)B5Nagy/J 003718 FVB-Tg(GadGFP)45704Swn/J 003773 STOCK Tg(CAG-ECFP)CK6Nagy/J 003920 STOCK Tg(CAG-Bgeo/GFP)21Lbe/J 003927 C57BL/6J-Tg(Sry-EGFP)92Ei/EiJ 004178 B6.129(Cg)-Tg(CAG-Bgeo/GFP)21Lbe/J 004218 B6.129(ICR)-Tg(CAG-ECFP)CK6Nagy/J 004353 C57BL/6-Tg(UBC-GFP)30Scha/J 004509 B6.FVB-Tg(Itgax-DTR/EGFP)57Lan/J 004512 C.FVB-Tg(Itgax-DTR/EGFP)57Lan/J 004654 B6;CBA-Tg(Pou5f1-EGFP)2Mnn/J 004690 B6;FVB-Tg(Pcp2-EGFP)2Yuza/J 005029 B6.Cg-Tg(Hlxb9-GFP)1Tmj/J 005070 C57BL/6-Tg(Csf1r-EGFP-NGFR/FKBP1A/TNFRSF6)2Bck/J 005104 STOCK Tg(tetO-HIST1H2BJ/GFP)47Efu/J 005244 B6.Cg-Tg(Krt1-15-EGFP)2Cot/J 005282 NOD/ShiLtJ-Tg(Ins1-EGFP/GH1)14Hara/HaraJ 005418 STOCK Tg(HIST1H2BB/EGFP)1Pa/J 005483 129-Tg(CAG-EYFP)7AC5Nagy/J 005515 FVB-Tg(ITGAM-DTR/EGFP)34Lan/J 005620 B6;D2-Tg(S100B-EYFP)1Wjt/J 005621 B6;D2-Tg(S100B-EGFP)1Wjt/J 005627 B6.Cg-Tg(Thy1-YFP/Syp)10Jrs/J 005630 B6.Cg-Tg(Thy1-EYFP)15Jrs/J 005699 STOCK Tg(tetO-Ipf1,EGFP)956.6Macd/J 005706 C57BL/6-Tg(tetO-CDK5R1/GFP)337Lht/J 005738 B6.FVB-Tg(tetO-EGFP,-Tgfbr2)8Mcle/J 005854 STOCK Tg(Cp-EGFP)25Gaia/J 006043 B6;SJL-Tg(Oxt/EGFP)AI03Wsy/J 006069 B6.Cg-Tg(HIST1H2BB/EGFP)1Pa/J 006129 STOCK Tg(Zp3-EGFP)1Dean/J 006361 B6.Cg-Tg(Sp7-tTA,tetO-EGFP/cre)1Amc/J 006567 C57BL/6-Tg(CAG-EGFP)131Osb/LeySopJ 006570 STOCK Smn1tm1Msd Tg(Hlxb9-GFP)1Tmj Tg(SMN2)89Ahmb/J 006614 B6;CBA-Tg(Thy1-CFP/COX8A)C1Lich/J 006617 B6;CBA-Tg(Thy1-CFP/COX8A)S2Lich/J 006665 B6;129P2-Olfr151tm13(rI7)Mom/MomJ 006850 STOCK Tg(CAG-Bgeo,-NOTCH1,-EGFP)1Lbe/J 006864 B6.Cg-Tg(Ins1-EGFP)1Hara/J 007075 CByJ.B6-Tg(CAG-EGFP)1Osb/J 007076 CByJ.B6-Tg(UBC-GFP)30Scha/J 007264 C57BL/6-Tg(Sry-EGFP)92Ei Tg(Sry)4Ei Chr YPOS/EiJ 007265 C57BL/6-Tg(Sry-EGFP)92Ei Chr YAKR/J/EiJ 007483 FVB.Cg-Tg(Tyr)3412ARpw Tg(Sry-EGFP)92Ei/EiJ 007484 B6.Cg-Tyrc-2J Tg(Tyr)3412ARpw Tg(Sry-EGFP)92Ei/EiJ 007567 C57BL/6J-Tg(Itgax-cre,-EGFP)4097Ach/J 007615 B6.Cg-Tg(Thy1-COP4/EYFP)9Gfng/J 007673 B6.Cg-Tg(Gad1-EGFP)3Gfng/J 007742 B6.Cg-Tg(Myh11-cre,-EGFP)2Mik/J 007857 C57BL/6J-Tg(Eno2-YFP/Cox8a)YRwb/J 007860 C57BL/6J-Tg(Eno2-YFP/Cox8a)ZRwb/J 007894 B6.Cg-Tg(Rgs4-EGFP)4Lvt/J 007940 B6.Cg-Tg(Thy1-CFP/COX8A)C1Lich/J 007967 B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J 007977 B6;CBA-Tg(Hf/Olfr16-GFP)7Mom/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 007986 B6;CBA-Tg(H*/Olfr16-GFP)11Mom/MomJ 007987 B6;CBA-Tg(H*/Olfr16-GFP)25Mom/MomJ 008004 B6;SJL-Tg(Thy1-ECFP/VAMP2)1Sud/J 008078 B6;129S4-Tcf3tm5Zhu/J 008126 B6.NOD-Tg(Cd4-EGFP)1Lt/J 008173 NOD.Cg-Tg(Ins1-EGFP)1Hara/QtngJ 008344 B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J 008450 FVB-Tg(CAG-luc,-GFP)L2G85Chco/J 008547 NOD.FVB-Tg(ITGAM-DTR/EGFP)34Lan/JdkJ 008549 NOD.FVB-Tg(Itgax-DTR/EGFP)57Lan/JdkJ 008579 STOCK Tg(PSCA-EGFP)1Witt/J 008694 NOD/ShiLt-Tg(Foxp3-EGFP/cre)1cJbs/J 008828 B6.Cg-Tg(Prdm1-EYFP)1Mnz/J 009159 B6;FVB-Tg(Cnp-EGFP/Rpl10a)JD368Htz/J 009593 C57BL/6J-Tg(Pomc-EGFP)1Low/J 010542 NOD.FVB-Tg(CAG-luc,-GFP)L2G85Chco/FathJ 010545 C.FVB-Tg(CAG-luc,-GFP)L2G85Chco/FathJ 010713 C57BL/6-Tg(tetO-GFP/tetX)5696Stl/J 010827 B6;C3-Tg(FOXJ1-EGFP)85Leo/J 011106 STOCK Tg(CAG-GFP*)1Hadj/J 012643 B6.Cg-Tg(Ly6a-EGFP)G5Dzk/J 013134 B6.Cg-Tg(Col1a1*2.3-GFP)1Rowe/J 013233 NOD.B6-Tg(Itgax-cre,-EGFP)4097Ach/J 014135 B6.Cg-Tg(Fos/EGFP)1-3Brth/J 014548 B6.Cg-Tg(Slc32a1-COP4*H134R/EYFP)8Gfng/J 017466 B6.Cg-Tg(Col1a1*3.6-Topaz)2Rowe/J 017467 C57BL/6-Tg(Dmp1-Topaz)1Ikal/J 017468 STOCK Tg(Col1a1*3.6-Cyan)2Rowe/J 017469 C57BL/6-Tg(BGLAP-Topaz)1Rowe/J 017472 STOCK Tg(Acp5-CFP,Ibsp-YFP,Dmp1-RFP)1Pmay/J 017484 FVB-Tg(JPH3-GFP,-JPH3*)GXwy/J 017495 B6;129S7-Crim1tm1(GFP/cre/ERT2)Pzg/J 017592 B6;129S-Sox2tm2Hoch/J 017619 NOD.Cg-Prkdcscid Tg(CAG-EGFP)1Osb/KupwJ 017952 STOCK Tg(Isl1-EGFP*)1Slp/J 018067 FVB-Tg(Prism)1849Htz/J 018068 FVB-Tg(Prism)1989Htz/J 018071 FVB-Tg(Prism)1861Htz/J 018322 STOCK Tg(Cp-EGFP)25Gaia/ReyaJ 018548 FVB/N-Tg(GFAP-Cadm1/EGFP)42Oje/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.