Stock Number: 008477 Strain Name: STOCK Tg(RARE-Hspa1b/lacZ)12Jrt/J

Allele: Generic LacZ Protocol Name: Generic LacZ Method: Standard PCR Version: 1.1 Created: 31-July -2000 Updated: 18-January -2012

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 315 bp Internal positive control = 210 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 oIMR0040 - CGT GGC CTG ATT CAT TCC - Transgene Reverse oIMR3054 - ATC CTC TGC ATG GTC AGG TC - Transgene Forward oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 10 X AB PCR BufferII 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 oIMR3054 0.30 0.50 uM 0.30 20 uM oIMR0040 0.30 0.50 uM 0.30 20 ul oIMR8744 0.30 0.50 ul 0.30 20 uM oIMR8745 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 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 20 sec - 3 64 30 sec* * -0.5 C per cycle 4 72 35 sec repeat steps 2-4 for 12 cycles 5 94 20 sec - 6 58 30 sec - 7 72 35 sec repeat steps 5-7 for 25 cycles 8 72 2 min - 9 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002193 C57BL/6J-Tg(MTn-lacZ)204Bri/J 002621 B6;SJL-Tg(tetop-lacZ)2Mam/J 002754 C57BL/6-Tg(LacZpl)60Vij/J 002856 FVB/N-Tg(TIE2-lacZ)182Sato/J 002865 B6CBA-Tg(Wnt1-lacZ)206Amc/J 002981 DBA/2-Tg(xstpx-lacZ)36And/J 002982 B6.Cg-Tg(xstpx-lacZ)32And/J 003139 B6.Cg-Tg(DBHn-lacZ)8Rpk/J 003299 B6;SWJ-Tg(TIMP3-lacZ)7Jeb/J 003315 FVB/N-Tg(tetORo1-lacZ)3Conk/J 003487 FVB.Cg-Tg(XGFAP-lacZ)3Mes/J 003754 B6.129S4-Shroom3Gt(ROSA53)Sor/J 004141 B6;CBA-Tg(UAS-lacZ)65Rth/J 004623 STOCK Tg(Fos-lacZ)34Efu/J 005317 B6.Cg-Tg(BAT-lacZ)3Picc/J 005493 STOCK Tg(Tek-rtTA,TRE-lacZ)1425Tpr/J 005707 STOCK Rag1tm1Mom Tg(TIE2-lacZ)182Sato/J 005728 STOCK Tg(tetO-Ipf1,lacZ)958.1Macd/J 005814 B6.129P2-Grm1tm1Dgen/J 006229 B6.Cg-Tg(DRE-lacZ)2Gswz/J 006465 B6;CBA-Tg(CAG-lacZ-WGA)330Bbm/J 006470 B6;129S-Hopxtm1Eno/J 006477 B6.Cg-Tg(CAG-lacZ-WGA)330Bbm/J 006578 STOCK Myoz2tm1Eno/J 006613 STOCK Tg(CAG-Bgeo,-Tle1,-ALPP)1Lbe/J 006665 B6;129P2-Olfr151tm13(rI7)Mom/MomJ 006672 B6;CBA-Tg(Olfr16*,taulacZ)7Mom/MomJ 006686 STOCK Olfr6tm1Mom/MomJ 006793 B6;CBA-Tg(P-taulacZ)13Mom/MomJ 007925 STOCK En2tm5.1Alj/J 008344 B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J 008477 STOCK Tg(RARE-Hspa1b/lacZ)12Jrt/J 008618 B6;A-Tg(OPN1LW-lacZ)1Nat/J 012335 STOCK Hprttm50(Ple55-lacZ)Ems/Mmjax 012353 STOCK Hprttm65(Ple53-lacZ)Ems/Mmjax 013144 B6.129P2-Aim2Gt(CSG445)Byg/J 018627 B6;SJL-Tg(Myl1-lacZ)1Ibdml/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.