Stock Number: 009583 Strain Name: B6(129S4)-Et(cre/ERT2)1957Rdav/J

Allele: Gt(ROSA)26Sortm1Sor Protocol Name: Gt(ROSA)26Sortm1sor STD Method: Robotic STD Version: 4.0 Created: 22-September-2010 Updated: 14-May -2012

Expected Results: Mutant = 340 bp Heterozygote = 340 bp and 650 bp Wild type = 650 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 oIMR8052 - GCG AAG AGT TTG TCC TCA ACC - - oIMR8545 - AAA GTC GCT CTG AGT TGT TAT - - oIMR8546 - GGA GCG GGA GAA ATG GAT ATG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 5.76 - 5.76 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00 33.3 X PM 105 0.36 1.00 X 0.36

Cycling  Step # Temp °C Time Note 1 94 3 min 65-1 2 94 30 sec - 3 65 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 003309 B6;129S4-Gt(ROSA)26Sortm1Sor/J 003310 129S-Gt(ROSA)26Sortm1Sor/J 006331 STOCK Gt(ROSA)26Sortm1(DTA)Jpmb/J 006911 B6;129-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm2(tetO-Pou5f1)Jae/J 006953 STOCK Notch1tm3(cre)Rko/J 006965 B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae/J 007670 B6;129S4-Gt(ROSA)26Sortm3(phiC31*)Sor/J 007689 129S4/SvJaeSor-Gt(ROSA)26Sortm4(attB/attP)Sor/J 007743 B6.129S4-Gt(ROSA)26Sortm3(phiC31*)Sor/J 007844 129S4/SvJae-Gt(ROSA)26Sortm2(FLP*)Sor/J 008706 B6.129P2(Cg)-Hprttm4(Ple88-EGFP)Ems/Mmjax 008707 B6.129P2(Cg)-Hprttm7(Ple185-EGFP)Ems/Mmjax 008708 B6.129P2(Cg)-Hprttm8(Ple151-EGFP)Ems/Mmjax 008709 B6.129P2(129S4)-Hprttm9(Ple178-EGFP/cre)Ems/Mmjax 008710 B6.129P2(129S4)-Hprttm10(Ple162-EGFP/cre)Ems/Mmjax 008876 STOCK Hprttm11(Ple176-EGFP/cre)Ems/Mmjax 008877 B6.129P2(129S4)-Hprttm12(Ple177-EGFP/cre)Ems/Mmjax 008883 B6;129-Gt(ROSA)26Sortm1(SNCA*A53T)Djmo/TmdJ 008886 B6;129-Gt(ROSA)26Sortm3(SNCA*E46K)Djmo/TmdJ 008889 B6;129-Gt(ROSA)26Sortm2(SNCA*119)Djmo/TmdJ 009427 FVB.129S4(B6)-Gt(ROSA)26Sortm1Sor/J 009573 B6(129S4)-Et(cre/ERT2)4Rdav/J 009577 B6(129S4)-Et(cre/ERT2)296Rdav/J 009579 B6(129S4)-Et(cre/ERT2)837Rdav/J 009581 B6(129S4)-Et(cre/ERT2)1642Rdav/J 009582 B6(129S4)-Et(cre/ERT2)1645Rdav/J 009583 B6(129S4)-Et(cre/ERT2)1957Rdav/J 009584 B6(129S4)-Et(cre/ERT2)2007Rdav/J 009585 B6(129S4)-Et(cre/ERT2)2047Rdav/J 009586 B6(129S4)-Et(icre)754Rdav/J 009587 B6(129S4)-Et(icre)1402Rdav/J 009588 B6(129S4)-Et(icre)1470Rdav/J 009589 B6(129S4)-Et(icre)1555Rdav/J 011013 STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm5(tetO-Pou5f1,-Klf4,-Myc)Jae/J 012580 B6.129P2(Cg)-Hprttm59(Ple123-lacZ)Ems/Mmjax 012581 B6.129P2(Cg)-Hprttm62(Ple153-lacZ)Ems/Mmjax 012583 B6.129P2(Cg)-Hprttm68(Ple127-lacZ)Ems/Mmjax 012584 STOCK Hprttm69(Ple134-lacZ)Ems/Mmjax 012687 B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J 012689 B6(129S4)-Et(icre/ERT2)14163Rdav/J 014602 B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-mCherry)Eggn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.