Stock Number: 002297 Strain Name: B6SJL-Tg(SOD1)2Gur/J

Allele: Tg(SOD) Protocol Name: Tg(SOD1) Method: Standard PCR Version: 1.1 Created: 09-April -2003 Updated: 20-February -2013

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 236 bp Internal positive standard = 324 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 oIMR0113 - CAT CAG CCC TAA TCC ATC TGA - Transgene oIMR0114 - CGC GAC TAA CAA TCA AAG TGA - 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 3.52 - 3.52 10 X AB PCR BufferII 0.84 0.70 X 0.84 25 mM MgCl2 0.48 1.00 mM 0.48 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR7338 0.45 0.75 uM 0.45 20 uM oIMR7339 0.45 0.75 uM 0.45 20 uM oIMR0113 0.80 1.33 uM 0.80 20 uM oIMR0114 0.80 1.33 uM 0.80 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.04 0.02 U/ul 0.04 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 95 3 min - 2 95 30 sec - 3 60 30 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 002297 B6SJL-Tg(SOD1)2Gur/J 002298 B6.Cg-Tg(SOD1)2Gur/J 002299 B6.Cg-Tg(SOD1*G93A)dl1Gur/J 002300 B6SJL-Tg(SOD1*G93A)dl1Gur/J 002628 C57BL/6-Tg(SOD1)10Cje/J 002629 C57BL/6-Tg(SOD1)3Cje/J 002726 B6SJL-Tg(SOD1*G93A)1Gur/J 004435 B6.Cg-Tg(SOD1*G93A)1Gur/J 008248 B6.Cg-Tg(SOD1*G85R)148Dwc/J 008342 B6.Cg-Tg(SOD1*G37R)42Dpr/J 013199 FVB.Cg-Tg(SOD1*G93A)1Gur/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.