Stock Number: 006883 Strain Name: B6.Cg-Ldlrtm1Her Sod2tm1Leb/J

Allele: Sod2tm1Leb Protocol Name: Human HPRT Melt Curve Analysis Method: Melt Curve Analysis Version: 2.1 Created: 17-September-2008 Updated: 09-November -2009

Expected Results: Mut Area = 4 Mut Tm = 91 +/- 1 WT Area = 8 WT Tm = 82 +/- 1

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0781 - TGT TCT CCT CTT CCT CAT CTC C - Mutant oIMR0782 - ACC CTT TCC AAA TCC TCA GC - Mutant oIMR0878 - TGA ACC AGT TGT GTT GTC AGG - Wild type oIMR0888 - TCC ATC ACT GGT CAC TAG CC - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.00 - 2.00 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR0781 0.80 1.33 uM 0.80 20 uM oIMR0782 0.80 1.33 uM 0.80 20 uM oIMR0878 0.20 0.33 uM 0.20 20 uM oIMR0888 0.20 0.33 uM 0.20 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 64 1 min - 4 72 30 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 002972 B6;129S7-Sod1tm1Leb/J 002973 B6.129S7-Sod2tm1Leb/J 003283 B6;129S7-Fshbtm1Zuk/J 003783 B6.129S7-Bmp7tm1Kry/J 006883 B6.Cg-Ldlrtm1Her Sod2tm1Leb/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.