Stock Number: 004155 Strain Name: B6.129S2-Alox5tm1Fun/J

Allele: Neo Protocol Name: Generic Neo Quantitative PCR-QPCR- 1.2 Method: QPCR Version: 1.0 Created: 15-June -2009 Updated: 24-September-2009

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1080 - CGG CTG CAT ACG CTT GAT C - Transgene oIMR1081 - CGA CAA GAC CGG CTT CCA T - Transgene oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse TmoIMR0001 6-FAM CAT CGC ATC GAG CGA GCA CGT ACT TAMRA Transgene TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 Probe

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X Universal Master Mix 10.00 0.80 X 10.00 40 uM oIMR1080 0.25 0.40 uM 0.25 40 uM oIMR1081 0.25 0.40 uM 0.25 40 uM oIMR1544 0.25 0.40 uM 0.25 40 uM oIMR3580 0.25 0.40 uM 0.25 5 uM TmoIMR0001 0.75 0.15 uM 0.75 5 uM TmoIMR0107 0.75 0.15 uM 0.75 ddH2O 2.50 - 2.50 DNA 10.00 - 10.00

Cycling  Step # Temp °C Time Note 1 50 2 min - 2 95 10 min - 3 95 15 sec - 4 60 1 min repeat steps 3-4 for 40 cycles

This genotyping protocol is used for the following strains:   Stock Number Strain Name 004155 B6.129S2-Alox5tm1Fun/J 006830 129-Dysftm1Kcam/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.