Stock Number: 004632 Strain Name: B6.Cg-Tg(GFAP-APOE*2)14Hol Apoetm1Unc/J

Allele: Tg(GFAP-APOE*2)14Hol Protocol Name: Tg(GFAP-APOE) Method: Standard PCR Version: 1.1 Created: 26-November -2003 Updated: 24-September-2009

Expected Results: TG = ~450bp internal control = 324 bp

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1940 - CCA GGG GGT GTT GCC AGG GGC ACC - Transgene oIMR1941 - TCC AGT TCC GAT TTG TAG GCC TTC AAC TCC - Transgene oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control 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 oIMR1940 0.60 1.00 uM 0.60 20 uM oIMR1941 0.60 1.00 uM 0.60 20 uM oIMR7338 0.30 0.50 uM 0.30 20 uM oIMR7339 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 63 45 sec - 4 72 45 sec 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 004631 B6.Cg-Tg(GFAP-APOE*4)1Hol Apoetm1Unc/J 004632 B6.Cg-Tg(GFAP-APOE*2)14Hol Apoetm1Unc/J 004633 B6.Cg-Tg(GFAP-APOE*3)37Hol Apoetm1Unc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.