Stock Number: 002207 Strain Name: B6.129S7-Ldlrtm1Her/J

Allele: Ldlrtm1Her Protocol Name: Ldlr tm1Her Method: Standard PCR Version: 4.0 Created: 23-December -2010 Updated: 23-December -2010

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 - Mutant 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) 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 ddH2O 4.46 - 4.46 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 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 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

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 7.06 - 7.06 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 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 KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 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-Lrpap1tm1Her Ldlrtm1Her/J 003000 B6;129S-Ldlrtm1Her Apobtm2Sgy/J 004192 STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J 006580 B6.Cg-Ins2Akita Ldlrtm1Her/J 006877 B6.Cg-Ldlrtm1Her Tg(H2-K-AKR1B1)1Tj/J 006883 B6.129S7-Ldlrtm1Her Sod2tm1Leb/J 006906 B6.Cg-Lepob Ldlrtm1Her/J 006952 B6.129-Akt2tm1.1Mbb Ldlrtm1Her/J 007068 D2.129S7(B6)-Ldlrtm1Her/J 007070 AK.129S7(B6)-Ldlrtm1Her/J 012845 CBy.129S7(B6)-Ldlrtm1Her/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  None found.