Stock Number: 008883 Strain Name: B6;129-Gt(ROSA)26Sortm1(SNCA*A53T)Djmo/TmdJ

Allele: Gt(ROSA)26Sortm1Sor Protocol Name: Gt(ROSA)26Sortm1sor STD Method: Standard PCR Version: 1.1 Created: 07-July -2008 Updated: 10-November -2009

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 - Mutant Reverse oIMR8545 - AAA GTC GCT CTG AGT TGT TAT - Common oIMR8546 - GGA GCG GGA GAA ATG GAT ATG - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR8545 0.60 1.00 uM 0.60 20 uM oIMR8546 0.60 1.00 uM 0.60 20 uM oIMR8052 0.60 1.00 uM 0.60 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 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 003474 B6.129S4-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 008706 B6.129P2-Hprt1tm4(Ple88-EGFP)Ems/J 008707 B6.129P2-Hprt1tm7(Ple185-EGFP)Ems/J 008708 B6.129P2-Hprt1tm8(Ple151-EGFP)Ems/J 008709 B6.129P2-Hprt1tm9(Ple178-EGFP/cre)Ems/J 008710 B6.129P2-Hprt1tm10(Ple162-EGFP/cre)Ems/J 008875 B6.129P2-Lgr5tm1(cre/ESR1)Cle/J 008876 B6;129-Hprt1tm11(Ple176-EGFP/cre)Ems/J 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 009574 B6(129S4)-Et(cre/ERT2)21Rdav/J 009575 B6(129S4)-Et(cre/ERT2)119Rdav/J 009576 B6(129S4)-Et(cre/ERT2)278Rdav/J 009577 B6(129S4)-Et(cre/ERT2)296Rdav/J 009579 B6(129S4)-Et(cre/ERT2)837Rdav/J 009580 B6(129S4)-Et(cre/ERT2)1382Rdav/J 009581 B6(129S4)-Et(cre/ERT2)1642Rdav/J 009583 B6(129S4)-Et(cre/ERT2)1957Rdav/J 009584 B6(129S4)-Et(cre/ERT2)2007Rdav/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 009608 B6.Cg-Hprt1tm36(Ple162-EGFP)Ems/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.