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

Allele: Tg(SOD1*G93A)1Gur Protocol Name: Tg(SOD) Method: Melt Curve Analysis Version: 2.1 Created: 16-September-2008 Updated: 24-September-2009

Notes: Melting curve analysis is done using a Roche Light Cycler 480. This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene Tm = 83°C Internal positive control Tm = 80°C

Gel Image

Image Image file View Full Size

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) 2 X SYBR Green Master Mix (Roche) 5.20 0.87 X 5.20 20 uM oIMR7338 0.60 1.00 uM 0.60 20 uM oIMR7339 0.60 1.00 uM 0.60 20 uM oIMR0113 0.80 1.33 uM 0.80 20 uM oIMR0114 0.80 1.33 uM 0.80 PBND 2.00 - 2.00 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 10 min - 2 94 30 sec - 3 65 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 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.