Stock Number: 008782 Strain Name: FVB.Cg-Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*A111G)588Ahmb/J

Allele: Tg(SMN2*A111G)588Ahmb/J Protocol Name: Tg(SMN2*A111G) Method: Standard PCR Version: 1.0 Created: 12-May -2009 Updated: 07-May -2010

Notes: May see doublet w/Tg band.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 8022 - TGG AGT TCG AGA CGA GGC CTA AGC - Transgene Forward oIMR8251 - AGT AGA TCG GAC AGA TTT TGC T - Transgene Reverse oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) DNA 2.00 - 2.00 ddH2O 3.16 - 3.16 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM 8022 0.90 1.50 uM 0.90 20 ul oIMR8744 0.09 0.15 ul 0.09 20 uM oIMR8745 0.09 0.15 uM 0.09 10 X High Fidelity Buffer 1.20 1.00 X 1.20 5 U/ul Platinum Taq Pol. 0.08 0.03 U/ul 0.08 5 M Betaine 1.66 0.69 M 1.66 20 uM oIMR8251 0.90 1.50 uM 0.90

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30sec - 3 59 1 min Go to step 2-4, 35 times 4 72 1 min - 5 72 2 min - 6 10 hold -

This genotyping protocol is used for the following strains:   Stock Number Strain Name 008782 FVB.Cg-Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*A111G)588Ahmb/J 009134 FVB.Cg-Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*A111G)591Ahmb/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.