Stock Number: 002726 Strain Name: B6SJL-Tg(SOD1*G93A)1Gur/J

Allele: Sod TgN Copy Number Protocol Name: Sod TgN Copy Number Method: QPCR Version: 2.0 Created: 05-October -2009 Updated: 05-October -2009

Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/Quencher combination.  The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1544 - CAC GTG GGC TCC AGC ATT - - oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - - oIMR9665 - GGG AAG CTG TTG TCC CAA G - - oIMR9666 - CAA GGG GAG GTA AAA GAG AGC - - TmoIMR0105 Cy5 CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 2 - TmoIMR0147 6-FAM CTG CAT CTG GTT CTT GCA AAA CAC CA Black Hole Quencher 1 -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X Universal Master Mix 6.25 1.00 X 6.25 40 uM oIMR9665 0.13 0.42 uM 0.13 40 uM oIMR9666 0.13 0.42 uM 0.13 40 uM oIMR1544 0.13 0.42 uM 0.13 40 uM oIMR3580 0.13 0.42 uM 0.13 5 uM TmoIMR0105 0.38 0.15 uM 0.38 5 uM TmoIMR0147 0.38 0.15 uM 0.38 DNA 5.00 - 5.00 ddH2O 0.00 - 0.00

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 002726 B6SJL-Tg(SOD1*G93A)1Gur/J 004435 B6.Cg-Tg(SOD1*G93A)1Gur/J 008229 B6.Cg-Tg(SOD1*G37R)29Dpr/J 008248 B6.Cg-Tg(SOD1*G85R)148Dwc/J 008342 B6.Cg-Tg(SOD1*G37R)42Dpr/J 308342 B6.Cg-Tg(SOD1*G37R)42Dpr/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.