Stock Number: 008228 Strain Name: C3.129S7(B6)-Ifngtm1Ts/J

Allele: Ifngtm1Ts Protocol Name: Ifngtm1Ts Method: Melt Curve Analysis Version: 9.0 Created: 17-June -2011 Updated: 20-May -2013

Expected Results: Mutant = ~500 bp Heterozygote = ~210 bp and ~500 bp Wild type = ~210 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 oIMR6218 - CCT TCT ATC GCC TTC TTG ACG - Mutant Forward oIMR8284 - AGA AGT AAG TGG AAG GGC CCA GAA G - Wild type Forward oIMR8285 - AGG GAA ACT GGG AGA GGA GAA ATA T - Common

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 20 uM oIMR8285 0.60 1.00 uM 0.60 20 uM oIMR8284 0.20 0.33 uM 0.20 20 uM oIMR6218 0.60 1.00 uM 0.60 ddH2O 2.60 - 2.60 DNA 2.00 - 2.00 2 X KAPA SYBR Fast 6.00 1.00 X 6.00

Cycling  Step # Temp °C Time Note 1 95 3 min KF 63 2 95 10sec - 3 63 45sec *35 cycles 4 72 3 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002286 C.129S7(B6)-Ifngtm1Ts/J 002287 B6.129S7-Ifngtm1Ts/J 002575 NOD.129S7(B6)-Ifngtm1Ts/DvsJ 008228 C3.129S7(B6)-Ifngtm1Ts/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.