Stock Number: 006665 Strain Name: B6;129P2-Olfr151tm13(rI7)Mom/MomJ

Allele: Generic GFP Protocol Name: Fluorescent Proteins (Generic GFP) Method: Standard PCR Version: 1.1 Created: 10-August -2001 Updated: 24-September-2009

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 - GFP/YFP/CFP oIMR1416 - TCC TTG AAG AAG ATG GTG CG - GFP/YFP/CFP 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) ddH2O 3.39 - 3.39 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 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 5 U/ul Taq DNA Polymerase 0.03 0.01 U/ul 0.03 DNA 2.00 - 2.00

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 005670 B6.Cg-Gt(ROSA)26Sortm1(rtTA,EGFP)Nagy/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 006582 B6.129S4-Park2tm1Shn/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 006686 STOCK Olfr6tm1Mom/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 007567 C57BL/6J-Tg(Itgax-cre,-EGFP)4097Ach/J 007572 B6.129P2(Cg)-Rorctm2Litt/J 007673 B6.Cg-Tg(Gad1-EGFP)3Gfng/J 007687 BKa.Cg-Sox17tm1Sjm Ptprcb Thy1a/J 007742 B6.Cg-Tg(Myh11-cre,-EGFP)2Mik/J 007843 B6;129S4-Efnb2tm2Sor/J 007894 B6.Cg-Tg(Rgs4-EGFP)4Lvt/J 007901 B6.Cg-Tg(Thy1-Brainbow1.0)HLich/J 007910 B6;CBA-Tg(Thy1-Brainbow1.0)LLich/J 007921 B6.Cg-Tg(Thy1-Brainbow2.1)RLich/J 007940 B6.Cg-Tg(Thy1-CFP/COX8A)C1Lich/J 007967 B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J 008004 B6;SJL-Tg(Thy1-ECFP/VAMP2)1Sud/J 008078 B6;129S4-Tcf3tm5Zhu/J 008126 B6.NOD-Tg(Cd4-EGFP)1Lt/J 008295 B6;129-Syt9tm1Sud/J 008344 B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J 008694 NOD/ShiLt-Tg(Foxp3-EGFP/cre)1Jbs/J 905341 Nestin GFP 905342 Nestin.GFP on C57BL/6 Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.