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

Allele: Generic GFP Protocol Name: Fluorescent Proteins (Generic GFP) Method: Melt Curve Analysis Version: 3.0 Created: 04-June -2009 Updated: 24-September-2009

Notes: Melting curve analysis is done using a Roche Light Cycler 480. This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene Tm = 89.5°C +/- 1.0°C Internal positive control Tm = 79.4°C +/- 1.0°C

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0872 - AAG TTC ATC TGC ACC ACC G - - oIMR3999 - TGC TCA GGT AGT GGT TGT CG - - 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 2.00 - 2.00 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR0872 0.80 1.33 uM 0.80 20 uM oIMR3999 0.80 1.33 uM 0.80 20 uM oIMR7338 0.20 0.33 uM 0.20 20 uM oIMR7339 0.20 0.33 uM 0.20 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 - 2 94 15 sec - 3 68 1 min - 4 72 30 sec 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 003920 STOCK Tg(CAG-Bgeo/GFP)21Lbe/J 004178 B6.129(Cg)-Tg(CAG-Bgeo/GFP)21Lbe/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 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 005282 NOD/ShiLtJ-Tg(Ins1-EGFP/GH1)14Hara/HaraJ 005418 STOCK Tg(HIST1H2BB/EGFP)1Pa/J 005670 B6.Cg-Gt(ROSA)26Sortm1(rtTA,EGFP)Nagy/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 006000 B6.FVB-Tg(ITGAM-DTR/EGFP)34Lan/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 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 007075 CByJ.B6-Tg(CAG-EGFP)1Osb/J 007264 C57BL/6-Tg(Sry-EGFP)92Ei Tg(Sry)4Ei Chr YPOS/EiJ 007567 C57BL/6J-Tg(Itgax-cre,-EGFP)4097Ach/J 007572 B6.129P2(Cg)-Rorctm2Litt/J 007673 B6.Cg-Tg(Gad1-EGFP)3Gfng/J 007788 STOCK Tg(Thy1-EGFP)MJrs/J 007857 C57BL/6J-Tg(Eno2-YFP/Cox8a)YRwb/J 007894 B6.Cg-Tg(Rgs4-EGFP)4Lvt/J 007901 B6.Cg-Tg(Thy1-Brainbow1.0)HLich/J 007919 B6.Cg-Tg(Thy1-EGFP)OJrs/GfngJ 007967 B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J 008126 B6.NOD-Tg(Cd4-EGFP)1Lt/J 008173 NOD.Cg-Tg(Ins1-EGFP)1Hara/QtngJ 008450 FVB-Tg(CAG-luc,-GFP)L2G85Chco/J 008694 NOD/ShiLt-Tg(Foxp3-EGFP/cre)1Jbs/J 009159 B6.FVB-Tg(Cnp-EGFP/Rpl10a)JD368Htz/J 009422 NOD.Cg-Tg(Itgax-EYFP)1Mnz/QtngJ 009593 C57BL/6J-Tg(Pomc-EGFP)1Low/J 010774 B6(Cg)-Calb2tm1(cre)Zjh/J 010776 B6(Cg)-Lhx6tm1(cre/ERT2)Zjh/J 010777 B6(Cg)-Pvalbtm1(cre/ERT2)Zjh/J 370329 Pax7-1 370330 Pax7-2 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.