Stock Number: 006906 Strain Name: B6.Cg-Lepob Ldlrtm1Her/J

Allele: Ldlrtm1Her Protocol Name: Ldlr tm1Her Method: Standard PCR Version: 2.1 Created: 30-April -2004 Updated: 09-November -2009

Notes: Use Perfect Match© (Stratagene) added before Taq to delete non-specific bands. Expected Results: Mutant = 350 bp Heterozygote = 167 bp and 350 bp Wild type = 167 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 oIMR0092 - AAT CCA TCT TGT TCA ATG GCC GAT C - - oIMR3349 - CCA TAT GCA TCC CCA GTC TT - Common oIMR3350 - GCG ATG GAT ACA CTC ACT GC - Wild type

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) 0.90 0.75 X 0.90 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR0092 0.80 1.33 uM 0.80 20 uM oIMR3349 0.80 1.33 uM 0.80 20 uM oIMR3350 0.20 0.33 uM 0.20 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 1 U/ul Perfect Match (Stratagene) 0.06 0.01 U/ul 0.06 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 30sec - 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 002077 B6;129S7-Ldlrtm1Her/J 002207 B6.129S7-Ldlrtm1Her/J 002245 B6;129-Apoetm1Unc Ldlrtm1Her/J 002246 B6.129-Apoetm1Unc Ldlrtm1Her/J 002465 B6;129S-Ldlrtm1Her Lrpap1tm1Her/J 003000 B6;129S-Apobtm2Sgy Ldlrtm1Her/J 004192 STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J 005993 B6;129S6-Pcsk9tm1Jdh/J 006580 B6.Cg-Ins2Akita Ldlrtm1Her/J 006877 B6.Cg-Ldlrtm1Her Tg(H2-K-AKR1B1)1Tj/J 006883 B6.Cg-Ldlrtm1Her Sod2tm1Leb/J 006906 B6.Cg-Lepob Ldlrtm1Her/J 006952 B6.Cg-Akt2tm1.1Mbb Ldlrtm1Her/J 006966 B6.Cg-Akt2tm1.1Mbb/J 007068 D2.129S7(B6)-Ldlrtm1Her/J 007070 AKR.129S7(B6)-Ldlrtm1Her/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.