Stock Number: 002878 Strain Name: B6;129-Apobtm1Sgy Apoetm1Unc/J

Allele: Apoetm1Unc Protocol Name: Apoetm1Unc Method: Standard PCR Version: 1.1 Created: 26-October -2000 Updated: 02-February -2010

Expected Results: Mutant = ~245 bp Heterozygote = ~155 bp and ~245 bp Wild type = ~155 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 oIMR0180 - GCC TAG CCG AGG GAG AGC CG - Common oIMR0181 - TGT GAC TTG GGA GCT CTG CAG C - Wild type Reverse oIMR0182 - GCC GCC CCG ACT GCA TCT - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.26 - 4.26 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 oIMR0180 0.30 0.50 uM 0.30 20 uM oIMR0181 0.30 0.50 uM 0.30 20 uM oIMR0182 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 30 sec - 3 68 40 sec - 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 002052 B6.129P2-Apoetm1Unc/J 002245 B6;129-Apoetm1Unc Ldlrtm1Her/J 002246 B6.129-Apoetm1Unc Ldlrtm1Her/J 002878 B6;129-Apobtm1Sgy Apoetm1Unc/J 002879 B6;129-Apobtm2Sgy Apoetm1Unc/J 004362 B6;129-Scarb1tm1Kri Apoetm1Unc/J 004631 B6.Cg-Tg(GFAP-APOE_i4)1Hol Apoetm1Unc/J 004632 B6.Cg-Tg(GFAP-APOE_i2)14Hol Apoetm1Unc/J 004633 B6.Cg-Tg(GFAP-APOE_i3)37Hol Apoetm1Unc/J 007067 D2.129P2(B6)-Apoetm1Unc/J 007069 STOCK Apoetm1Unc/J 007224 B6.Cg-Apoetm1Unc (D12Mit182-D12Mit2)/Pgn 008525 B6.Cg-Tg(Cyp21a1-Apoe)619Fet/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.